ObjectiveTo investigate the anti-tumor effect of agonist MnCl_2of a novel cyclic guanosine monophosphate-adenosine monophosphate synthase(c GAS)/stimulator of interferon genes(STING)pathway collaborated with tumor cell lysate(Lysate)and the neo-antigen 10K-Adpgk of mouse colon cancer MC38 cell line.MethodsBone marrow-derived dendritic cells(BMDCs)were extracted from mouse bone marrow and divided into three groups:PBS,1 μmol/L MnCl_2and 10 μmol/L MnCl_2,which were analyzed for the maturation by flow cytometry,determined for the concentration of IL-6 in supernatant by ELISA,and detected for the transcription levels of IL-6,IFN-α,IFN β and CXCL9 genes by q PCR.Mouse tumor model was established by using MC38 cell line.When the tumor volume reached 100 mm3,the mice were randomly divided into two groups for administration,PBS,Lysate,MnCl_2,10K-Adpgk,Lysate + MnCl_2group and Lysate +10K-Adpgk + MnCl_2combined treatment group,which were administered subcutaneously through the tail for 3 times,with each interval of 1 week,and measured for the tumor volume every 2 days.One week after the last dose,serum samples were collected and determined for the concentrations of IFNγ and TNFα by ELISA.The tumor and spleen were isolated.The proportions of tumor infiltrating T cells and T cells in peripheral blood mononuclear cells(PBMCs)and the ratio of T cells to memory T cells in spleen were detected by flow cytometry,and the proportion of antigen specific T cells in spleen was detected by ELISPOT.Results10 μmol/L MnCl_2stimulated the maturation of BMDCs and activated the subsequent immune process.The tumor volumes of mice in the combined treatment group were considerably smaller than those in PBS group,the contents of IFNγ and TNF-α in serum were higher than those in other groups,and the proportions of tumor infiltrating T cells,T cells in PBMCs and ratio of T cells to memory T cells in spleen were also significantly higher than those in PBS group.Combined therapy caused strong antigen-specific T cell immune response.ConclusionThe addition of the novel adjuvant MnCl_2significantly enhanced the treatment effect of tumor cell lysate and neo-antigen,which provided an experimental basis for the development of the combination tumor treatment method based on MnCl_2and tumor antigens.

Objective To investigate the clinical significance of the changes of hemodynamics and electrolyte during orthotopc liver transplantation.Methods We studied 9 patients undergoing elective orthotopic liver transplantation without venovenous bypass,drew out arterial blood for monitoring arterial blood gas tensions,Na~+,K~+,Ca~(2+) and observed changes in hemodynamics and electrolyte.Results In most patients,arterial pressure significantly decreased at the prophase of the anhepatic phase,and tended to normal range in other phases.The concentration of calcium maintained a low-level during the whole surgery.During the prophases of the anhepatic phase and neo-hepatic phase,there was a light hyperkalemia,and the concentration of natrium slightly increased since 30 minutes after the anhepatic phase and neohepatic phase,but insensibly exceeded normal range.With resuming of the stability of hemodynamics,these changes had followed.Conclusion We shoud give our attention to correct the occurrence of the hypocalcemia during the forepart of the anhepatic phase and neo hepatic phase,and care for the hyperkalemia 5 minutes after the block of liver blood purveyance and revascularization of liver.The lock of body capability shoud be recruit duly according to the lose of blood and hemacytometer changes.In order to prevent serious academia and maintain the stability of hemodynamics and electrolyte,vasoconstrictor shoud be used carefully during the anhepatic phase.

AIM:To evaluate the influence of scutellarin on the expression of vascular endothelial growth factor ( VEGF) in high glucose-treated human retinal pigment epithelial cell line ARPE-19 and to observe the effects of scutellarin on the protein expression of VEGF, p-ERK and VEGFR2 in the retinas of type II diabetic rats.METHODS: Cultured ARPE-19 cells were divided into normal control group, scutellarin group, high glucose group and high glucose+scutellarin group.The protein levels of VEGF, p-ERK and VEGFR2 were measured by Western blot.The VEGF release in ARPE-19 cells was detected by ELISA.Normal rats were randomly divided into normal control group and scutellarin group.Diabetic rat model was established by feeding with high-fat diet and injecting with streptozocin, and randomly divided into diabetes group and diabetes treated with scutellarin group.After 16 weeks, the eyes were removed.The morphological changes of the retinas were observed under light microscope with HE staining, and histopathological score was recorded.The expres-sion of VEGF in the retinas was observed by the method of immunohistochemistry.RESULTS:Compared with normal con-trol group, the protein levels of VEGF, p-ERK and VEGFR2 in the ARPE-19 cells decreased in scutellarin group, but in-creased in high glucose group.The histopathological score of the retinas showed significant difference among diabetes group, diabetes treated with scutellarin group and normal control group, and no significant difference between normal con-trol group and scutellarin group was observed.The expression of VEGF increased in diabetic group and was significantly higher than that in scutellarin treatment group (P<0.05).CONCLUSION:Scutellarin inhibits the increased protein le-vels of VEGF, p-ERK and VEGFR2 in ARPE-19 cells, and decreases the expression of VEGF in the retinas of diabetic rats.The suppression of the diabetic retinopathy development by scutellarin may be partly involved in the ERK/MAPK pathway.

Objective To investigate the clinical significance of fine needle aspiration biopsy and immunohistochemical markers in differential diagnosis of thyroid nodule.Methods One hundred and eighty-five cases of thyroid nodule were enrolled in the study.All cases received preoperative fine-needle aspiration biopsy and immunocytochemistry.Results The sensitivity,specificity,diagnostic accuracy of fine needle aspiration biopsy to detect diagnosis of thyroid nodules was 75.0％ (21/28),85.4％ (134/157),83.8％(155/185).The positive rates of p53 and galectin-3 immunodetection in discriminating benign from malignant thyroid nodules were 60.7％ (17/28),85.7％ (24/28),combined with p53 and galectin-3 immunodetection in discriminating benign from malignant thyroid nodules was 50.0％ (14/28).The positive rates of p53 and galectin-3 immunodetection in thyroid adenomas were 0,5.1％ (8/157),combined with p53 and galectin-3 immunodetection in thyroid adenomas was 1.3％ (2/157).The positive rates of p53 and galectin-3 immunodetection in malignant thyroid cytologic specimens,alone or combined were significantly higher than those in thyroid adenomas,there were significant difference(P ＜ 0.05).The diagnostic accuracy of fine needle aspiration biopsy with p53 and galectin-3 immunodetection,alone or combined,in discriminating benign from malignant thyroid nodules [94.1％(174/185),93.5％(173/185),91.4％(169/185)] were higher or equal to those in fine needle aspiration biopsy[83.8％(155/185)],there were significant differences (P ＜0.05).Conclusion Immunodetection of p53 and galectin-3 combined with fine needle aspiration biopsy in the differential diagnosis of thyroid nodules can raise the diagnostic accuracy.

Objective To identify differentially expressed proteins in keloid tissue and to explore the proteomic characteristics of keloid by utilizing capillary high-performance liquid chromatography (LC) and tandem mass spectrometry (MS).Methods Tissue specimens were obtained from the lesions of 5 patients with non-familial keloid and normal skin of 5 human controls.After extraction and in-solution digestion of proteins,the resultant peptides were separated and identified by LC-MS/MS.Label-free detection and quantitation of peptides were carried out by using the DeCyder MSTM software.Results A total of 570differentially expressed peptides were identified between non-familial keloid and normal tissue (all P ＜ 0.05),which correspond to 1221 proteins.A difference greater than 1.5 folds was observed in the expressions of 293 proteins,including 124 up-regulated and 169 down-regulated proteins in non-familial keloid tissue.Conclusion Differences exist in the expression of proteins between non-familial keloid and normal skin tissue.

AIM: To study the effect of sesamin on expression of inducible nitric oxide synthase(iNOS)and nitrotyrosine(NT)in rat liver tissue with metabolic syndromic hepatic steatosis. METHODS: The rat model of metabolic syndromic hepatic steatosis was induced by operation of two kidneys with one clip(2K1C)and high-fat. The rats taken from that successful model were randomly divided into model group and sesamin(120, 60, and 30 mg·kg~(-1)·d~(-1))groups. In addition, the sham-operated group was set up. The rats in treated group were given sesamin intragastrically everyday for 8 weeks. The levels of blood lipids(TC, TG and FFA)in serum were detected. The activity of SOD and MDA level in the liver homogenate were determined. The expressions of iNOS and NT proteins were detected by Western blotting analysis. The histopathological changes were observed by HE staining in the liver tissues. RESULTS: Compared to model groups, sesamin(120, 60 mg·kg~(-1))significantly inhibited the elevation of serum TC, TG, FFA, and MDA in liver homogenate(P<0.05), and increased the activity of SOD(P<0.05). It also decreased the protein expression of iNOS and NT(P<0.05), and ameliorated the degree of hepatic steatosis. CONCLUSION: Sesamin prevents and cures the metabolic syndromic hepatic steatosis. The mechanism is probably mediated through decreasing the protein expression of iNOS and NT, and alleviating the oxidative stress in addition to regulating the lipid metabolism.

Objective: To investigate pharmacokinetics of propofol during anhepatic period of orthotopic liver transplantation (OLT) in children. Methods: In 5 children undergoing OLT, 2 mg/kg propofol was injected intravenously at the beginning of induction and anhepatic periods respectively. The plasma concentration of propofol was measured by reverse-phase high performance liquid chromatography(RP-HPLC), and the pharmacokinetics character of propofol between anhepatic and inducing periods were compared. Results: Propofol had wide distribution and high clearance, and its plasma concentration-time curves were fitted to a three-compartment open model. Conclusion: There is no difference of propofol metabolism between anhepatic and inducing periods, suggesting that there must be notable extra-hepatic metabolism when propofol used in children undergoing OLT.

Objective To detect differentially expressed genes in prostate cancer by cDNA microarray. Methods Total RNA was isolated by one step protocol from prostate cancer (CaP) and from normal prostate,and Poly(A) +RNA was further purified through Oligitex mRNA Midi Kit.Then it was analysed for differentially expressed genes in CaP and normal prostate by cDNA microarray with 4 096 human genes. Results There were 341 differentially expressed genes between CaP and normal prostate,of which there were 128 down regulated and 213 up regulated ones for CaP. Among these genes,15 were the most significant with 6 down regulated and 9 up regulated. Conclusions cDNA microarray can be used effectively to find out differentially expressed genes in prostate cancer which is not regulated by any single gene.Many different kinds of genes are involved in the initiation and evolution of CaP carcinogenisis.

Objective To study the expression and significance of antiapoptosis facter-clusterin in prostate neoplasm. Methods Reverse transcriptive polymerase chain reaction(RT PCR) was used to examine the expression level of clusterin in prostate cancer tissues(3 cases),prostate cancer cell line PC3M and normal prostate tissues(3 cases). Results Compared with the internal marker gene ? actin , the expression level of clusterin in prostate cancer is much higher than that of normal prostate. Conclusions The high expression level of clusterin in prostate cancer indicates that clusterin may play an important role in the biological characteristics of prostate cancer through the antiapoptosis pathway.

Objective To investigate the expression and significance of T-bet in experimental autoimmune uveoretinitis (EAU). Methods EAU was induced in 24 Lewis rats by immunization with retinal S-antigen (50 ?g) and complete Freund′s adjuvant, and another 4 rats were the healthy control. The rats with EAU were executed 7, 12, 15, 21 days after immunization, respectively. Immunohistochemical single and double staining were performed using monoclonal antibodies of T-bet or CD4 on the ocular wholemounts and the consecutive sections of the eye and spleen from both 24 immunized Lewis rats and 4 normal controls. The positive cells were counted under the optic microscope and the data were analyzed by SPSS 11.0 statistic software. Results A few T-bet positive cells were observed in the normal ocular tissues and spleen. The expressions of T-bet in the iris, retina, and spleen increased 7 days after immunization, reached the peak at the 15th day, and decreased at the 21st day, which were higher than those in the control. Double staining on the consecutive sections revealed that most of the T-bet positive cells were positive for CD4 monoclonal antibody. Conclusion T-bet may affect the occurrence of EAU by activating Th1 cells.