1.The induction of tolerogenic dendritic cells by curcumin and its mechanism
Heng-Cheng ZHU ; Xiu-Heng LIU ; Tao TAO ;
Chinese Journal of Organ Transplantation 2005;0(11):-
Objective To observe the induction of tolerogenic dendritic cells (DCs) by curcumin (Cur) and its mechanism.Methods After immature DCs from bone marrow cells of Wistar rats were treated with different concentrations of Cur (0,10,20 and 30?mol/L) respectively,and then the DCs were tested by flow cytometry for the surface molecules expression.After the immature DCs were treated by 30?mol/L Cur with or without stimulation of LPS,endocytosis of DCs to dextran was tested by flow cytometry.The production of IL-12 in DC culture supernatant was determined by ELISA.The levels of NF-?B p65 and RelB translocation to the nucleus were investigated by Western- blot.The activity of NF-?B was detected by NF-?B-binding ELISA and luciferase reporter gene analy- sis.The ability of DCs to stimulate the proliferation of T cells from Lewis rats were analyzed by mixed leukocyte reactions (MLR).Results Cur suppressed LPS-indueed cell-surface expression of costimu- latory molecules (CD80,CD86 and CD40) in a dose-dependent manner.When Cur was used at a con- centration of 30?mol/L,there was no marked difference in the surface molecules expression of LPS- inducing DCs as compared with immature DCs.After DCs were induced by LPS (LPS group),the positive rate of FITC-Dextran uptake was (36.6?7.2)%,and the secretory amounts of IL-12 were (415.9?42.7) pg/ml.In DCs of LPS group,the intranuclear RelB and p65 were highly expressed and their DNA binding activity was 0.65?0.08 and 0.74?0.07 respectively.The luciferase activity of reporter gene in LPS group DCs was remarkably increased to 435% as compared with that in the controls.DCs in LPS group showed strong capacity to stimulate T cells proliferation.When DCs were treated with 30?mol/L Cur followed by induction with LPS (Cur+LPS group),the positive rate of Dextran uptake was (78.6?14.2)% and remarkably higher than in LPS group (P
3.Influence of different antigen retrieval on the immunohistochemistry.
Xin-lan LUO ; Xiu-ling CAI ; Yan-hui LIU ; Heng-guo ZHUANG ; Wei ZHANG
Chinese Journal of Pathology 2005;34(1):52-54
Antigens
;
analysis
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Autoantigens
;
analysis
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Breast Neoplasms
;
metabolism
;
Citrates
;
Female
;
Formaldehyde
;
Hot Temperature
;
Humans
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Hydrogen-Ion Concentration
;
Immunohistochemistry
;
methods
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Iodide Peroxidase
;
analysis
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Iron-Binding Proteins
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analysis
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Paraffin Embedding
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Receptors, Progesterone
;
analysis
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Thyroid Gland
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immunology
;
Tissue Fixation
4.Effects of pH9.0 Tris-EDTA and pH6.0 citrate on antigen retrieval.
Xin-Lan LUO ; Yan-Hui LIU ; Heng-Guo ZHUANG ; Xiu-Ling CAI ; Jie XU
Chinese Journal of Pathology 2005;34(11):749-751
Antigens
;
analysis
;
Cervical Intraepithelial Neoplasia
;
metabolism
;
Citric Acid
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Cyclin-Dependent Kinase Inhibitor p21
;
analysis
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Edetic Acid
;
Formaldehyde
;
Hot Temperature
;
Humans
;
Hydrogen-Ion Concentration
;
Immunohistochemistry
;
Intestinal Mucosa
;
immunology
;
Ki-67 Antigen
;
analysis
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Microwaves
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Palatine Tonsil
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metabolism
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Proto-Oncogene Proteins c-bcl-6
;
analysis
5.Isolation and Identification of Seven Symbiotic Bacteria from Local Entomopathogenic Nematodes
Xiu-Fen YANG ; Zheng LIU ; Ran ZHANG ; Huai-Wen YANG ; Jing-Jing YUAN ; Heng JIAN ;
Microbiology 1992;0(02):-
The symbiotic bacterium exists in the intestines of entomopathogenic nematodes and is a potential biological agent.Systematic classification of these bacteria is scarce in China.In this paper,seven strains of symbiotic bacteria from local entomopathogenic nematodes were identified by both observation of mor-phology,physiological,biochemical characteristics and sequence analysis of 16S rDNA fragments.
6.Prostate cancer cell vaccine transfected with 4-1BBL induces anti-tumor immunity in vitro.
You-lin KUANG ; Xiao-dong WENG ; Xiu-heng LIU ; Zhi-yuan CHEN ; Heng-cheng ZHU ; Bo-tao JIANG
National Journal of Andrology 2010;16(9):773-777
OBJECTIVETo explore the anti-tumor immunity in vitro induced by prostate cancer cell vaccine transfected with recombinant adenovirus encoding 4-1BBL in mice.
METHODSThe replication-deficient adenovirus AdEasy-1 system was used to construct recombinant adenovirus Ad-m4-1BBL and Ad-eGFP. The prostate cancer cell RM-1 of mice was transfected with Ad-m4-1BBL and Ad-eGFP, and treated with mitomycin (MMC) to produce TCV, TCV-Ad-eGFP and TCV-Ad-m4-1BBL, followed by co-culture with syngeneic murine spleen cells. Then the cytotoxic activity of the lymphocytes against RM-1 cells was analyzed with CCK-8 solution, and IL-2 and INF-gamma were detected by ELISA.
RESULTSThe 4-1BBL protein was highly expressed in the TCV-Ad-m4-1BBL of the 4-1BBL-transfected mice. TCV-Ad-m4-1BBL significantly increased the expressions of IL-2 ([180.24 +/- 2.22] pg/ml) and INF-gamma ([1512.46 +/- 23.64] pg/ml) as compared with TCV and TCV-Ad-eGFP (P < 0.05), and induced higher RM-1 cell specific cytotoxicity ([34.24 +/- 2.64]%) than the latter two ([9.82 +/- 1.48]%) and ([14.65 +/- 3. 21]%), (P < 0.05). But none of them exhibited significant cytotoxicity against hepatocellular carcinoma Hepal-6.
CONCLUSIONThe m4-1BBL-expressing prostate cancer cell vaccine can effectively induce anti-tumor immune responses.
4-1BB Ligand ; genetics ; immunology ; Animals ; Cancer Vaccines ; genetics ; immunology ; Cell Line, Tumor ; Coculture Techniques ; Cytotoxicity, Immunologic ; genetics ; Female ; Interleukin-2 ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Prostatic Neoplasms ; Transfection
7.Effects of 75 gram glucose oral tolerance test and standard meal test on insulin secretion function of islets of Langerhans and free fatty acid in patients with type 2 diabetes mellitus
Qi SUN ; Jing-Bo ZENG ; Kang YU ; Yu-Xiu LI ; Qiu-Ying LIU ; Wei QIN ; Qing-Rong PAN ; Heng WANG ;
Chinese Journal of General Practitioners 2005;0(07):-
Objective To observe the effects of 75 gram glucose oral tolerance test (75 g OGTT) and standard mixed meal test (SMMT) on insulin secretion function of the islets of Langerhans and plasma free fatty acid (FFA) in patients with type 2 diabetes mellitus.Methods Seventy-six patients with type 2 diabetes without using insulin and with no obvious complications were recruited for 75 g OGTT following overnight fasting on the first day and SMMT (bread 50 g,egg 50 g and milk 250 ml) on the 7th day.Blood specimens were collected from each patients before the tests and 30 min,60 min,120 min and 180 min after glucose or meal load to measure their levels of plasma glucose,serum insulin,C peptide,FFA and lipids (total cholesterol,triglyceride,high-density and low-density lipoprotein cholesterol).Results No difference in fasting plasma glucose,serum insulin,C peptide,FFA and lipids between 75 g OGTT and SMMT was found.Postprandial plasma glucose 30 min,60 min,120 min and 180 min after 75 g OGTT was significantly higher than that after SMMT,with (15.3?3.5) vs (9.9?3.4) mmol/L,(18.2?4.8) vs (12.8?4.0) mmol/L,(16.3?5.8) vs (12.2?4.9) mmol/L and (10.6?5.4) vs (9.5?4.5) mmol/L (F=28.1,P
8.Exploration of differential expressed genes involved in the development and progression of hepatocellular carcinoma using oligo microarray.
Xiu-feng LIU ; Rui-hua SHI ; Heng-jun GAO ; Hong ZHU ; Shu-kui QIN ; Xue-hao WANG
Chinese Journal of Hepatology 2006;14(11):824-827
OBJECTIVESTo analyze the differential expression genes (DEGs) among hepatocellular carcinoma (HCC), para-cancerous tissue (PCT) and normal liver tissue (NLT) and explore the target genes related to the development and progression of HCC.
METHODSThe total RNAs of matched HCC, PCT and NLT of HCC patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis and lab-on-chip. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of HCC and NLT, HCC and PCT were hybridized with Agilent oligo microarray (21,074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by SYBR Green I stained real time RT-PCR.
RESULTS(1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 420 up-regulated genes and 552 down-regulated genes among 2-fold DEGs, including DKK1 (dickkopf homolog 1) which was 5-fold up-regulated; (3) The results of real time RT-PCR, using beta-actin as an internal control, showed that the 2-Delta Ct values of DKK1 in HCC, PCT and NLT were 0.089 504, 0.007,65 and 0.000,631 respectively.
CONCLUSION(1) The high throughput and effective Agilent oligo microarray can screen novel therapy targeted genes by analyzing the DEGs in development and progression of HCC; (2) The development and progression of HCC is a complicated process involving multigenes and multiprocedures; (3) DKK1, as a novel gene, is involved in the development and progression of HCC and may be a new therapy target.
Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis ; RNA, Neoplasm ; genetics
10.Buried penis decreases nitric oxide synthase of the corpus cavernosum.
Wei-Min YU ; Fan CHENG ; Xiao-Bin ZHANG ; Xiu-Heng LIU ; Yue XIA ; Ming-Huan GE
National Journal of Andrology 2008;14(5):411-413
OBJECTIVETo explore the influence of buried penis on nitric oxide synthase (NOS) activity of the corpus cavernosum in rats.
METHODSThe experimental model of concealed penis was established by intra-pocket-suture of the root of the penis. Two hundred and forty rats were equally randomized into a 2, a 4 and a 6 months group, each further divided into a buried (n = 50), a sham operation (n = 15) and a normal subgroup (n = 15). The development of the corpus cavernosum was surveyed by measuring its weight and the ratio to the body weight, followed by determination of NOS activity in the corpus cavernosum by spectrophotometry.
RESULTSNo significant differences were found in the corpus cavernosum weight, the body weight and their ratio among the buried, sham operation and normal groups in any experimental stage (P > 0.05). Buried penis decreased NOS activity in the 4- and 6-month groups (P < 0.05 and P < 0.01) compared with the normal group, but effected no significant change in the 2-month group.
CONCLUSIONBuried penis decreases the NOS activity of the corpus cavernosum in a positively time-related manner, but with no significant influence on its appearance and weight.
Animals ; Disease Models, Animal ; Erectile Dysfunction ; enzymology ; physiopathology ; Male ; Nitric Oxide Synthase ; metabolism ; Penis ; abnormalities ; enzymology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry