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.

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.

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.

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 relationship between occurrence of unexplained recurrent spontaneous abortion(URSA) and immune function of T lymphocytes. Methods T lymphocyte subsets and CD + 4 T helper subsets (including T helper 1 and T helper 2 cells) in peripheral blood from 30 cases with URSA and 24 healthy women (controls) were detected by enzyme-linked immunospot assay (ELISPOT). Results The percentage of CD + 4 positive T lymphocytes in URSA group was higher than that in the controls (60% vs 45%, P

Objective Cloning of some specific genes related to the prostate cancer. Methods Specimen from a patient with prostate cancer and from a normal adult were studied by the improved mRNA differential display and the differentially expressed sequence-tags were cloned,sequenced and analized.Reverse transcriptive-polymerase chain reaction(RT-PCR) was used to examine the expression level of Clusterin in prostate cancer tissues,prostate cancer cell line and normal prostate tissues. Results Significant difference was observed between the two kinds of tissues in gene expression and seven differentially expressed sequence-tags were found through analysis in GenBank.Among them,five were proved to be new gene tags,one shared 100% homology with Clusterin,which was highly expressed in the prostate cancer tissue and the other one shared 97% homology with superoxide dismutase 1 (SOD1),which was highly expressed in the normal prostate tissue.It was proved by RT-PCR that to compare with the internal marker gene ?-actin,the expression level of clusterin in prostate cancer is much higher than that of normal prostate. Conclusions Five new gene tags,clusterin and SOD1 were found to be differentially expressed between tissues of prostate cancer and normal prostate and may play important roles in carcinogenesis and development of prostate cancer.

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 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.

Objective To investigate the expression of T cell receptor (TCR) V ?8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Interphotoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) method was used to isolate the CD4+ T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR V ?8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+ T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P