1.In situ observation of surface structures of cardiovascular endothelial cells with atomic force microscope
Tong YIN ; Jin LUO ; Yamin MA ; Xiaolong JI ; Yusheng ZHAO ; Shiwen WANG
Journal of Geriatric Cardiology 2009;6(3):178-181
Objective To observe the surface structures of cardiovascular endothelial cells in situ with atomic force microscope (AFM). Methods Fresh aorta and aortic valve were dissected from 10 healthy male New Zealand white rabbits. Before fixed in 1% formaldehyde, the fresh tissues were washed in the buffer phosphate solution. Under general microscope, the fixed aorta or valve was spread on the double side stick tape which had already been stuck on the glass slide. The intima of aorta or the aorta side of valve was towards upside. Then the specimen was dried under 37 degrees centigrade in an attemperator and was washed with pure water. After dried again, the specimen was loaded on the platform ofNanoScope llla AFM and was scanned in tapping mode with the scanning speed of 0.5 HZ. Results The surface structures of endothelial cell on the fixed and dried tissue could be obsserved clearly in situ with AFM. Aortic endothclial cells were large, branched and arranged sparsely and parallel to the direction of blood flow, whereas endothelial cells on aorta valve surface were small, less branched and arranged intensively and vertical to the direction of blood flow. When the scanning range was dwindled, granular ultra-structures could be observed on the surface of endothelial cells, and, as the scanning range was dwindled further, fissure and convolution could be seen on the surface of granules from aortic endothelial cells. Centre cavity and surrounding swelling volcano-like structure could be seen on the surface of granules from endothelial cells of aortic valve. Conclusions It's feasible to observe the surface ultra-structures of cardiovascular endothelial cells in situ with AFM and morphological information provided by A FM might be of clinical value in future histopathological diagnosis.
2.Analysis of Chemical Components and Antifungal Activity of Extraction from Conidia of Trichoderma viride LTR-2
Kai CHEN ; He-Tong YANG ; Ji-Shun LI ; Jin-Dong HU ; Guang-Zhi ZHANG ;
Microbiology 1992;0(03):-
To study the chemical components and the antifungal activity of extraction from conidia of Trichoderma viride LTR-2.The extraction were obtained by distilling with Methylene dichloride from conidia of Trichoderma viride LTR-2 cultured on wheat bran solid matrix.Antifungal activity were determined by mycelium growth method.The chemical components of the extraction were analysed by GC-MS,the relative components in the extraction were determined by area normalization.The extraction not only have broad-spectrum control,showed antibiosis against eleven different plant fungal pathogens in PDA dish,such as Rhizoctonia solani,Alternaria brassica,Verticillium dahliae,Macrophoma kawatsukai,Fusarium moniliforme,Botrytis cinerea,Rhizoctonia cerealis,Fusarium oxysporum f.sp.vasinfectum,Bipolaris sorokinana,Fusarium graminearum,Alternaria.mali,but also have high inhibitory effect,and had 89.3% suppressive rate to Rhizoctonia cerealis.About sixty components were separated and identified by GC-MS,majority components were Hydrocarbon,the number of the Hydrocarbon were fourty-three kinds.Ergosterol was the major chemical components of the extract,and has 41.90% content.Other components comprised:Ketone,Organic acid,Alcohol,Ene,et al.Conclusion:The extraction from conidia of Trichoderma viride LTR-2 have antifungal activity.The extration comprised 2H-Pyran-2-one,5,6-dihydro-6-pentyl,it has 2.35% content.reference others literature,2H-Pyran-2-one,5,6-dihydro-6-pentyl may be the suppressive component of the extration.
4.Comparison of surface marker of monocyte-derived dendritic cells between cord blood and adult peripheral blood
ji-hong, QIAN ; tong-xin, CHEN ; xi, WANG ; ying-ying, JIN ; jian-xing, ZHU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(10):-
0.05).The percentage of CD40 positive cells in CBMC-derived DC was lower than that in PBMC-derived DC[(34.80?7.77)% vs(54.37?9.57)%,P
5.Multi-epitope DNA vaccines against avian influenza in chickens.
Jin-Mei PENG ; Guang-Zhi TONG ; Yun-Feng WANG ; Hua-Ji QIU
Chinese Journal of Biotechnology 2003;19(5):623-627
Multiple epitopes from one or more viruses can be lined up and co-expressed in one vector to generate multi-epitopes DNA vaccines. In the study, four recombinant plasmids were constructed based on HA and NP gene of avian influenza virus (AIV) (H5N1): (1) pIRES/HA, carrying the complete HA gene; (2) pIRES/tHA, carrying a truncated HA gene fragment of major neutralizing antigenic epitopes; (3) pIRES/tHA-NPep, in which three CTL epitopes of NP gene of AIV were fused to the truncated HA from the C-terminal; and (4) pIRES/tHA-NPep-IFN-gamma, which was constructed by replacing neo gene in pIRES/ tHA-NPep with IFN-y of chicken. Fifty five SPF chickens were randomly divided into five groups and immunized with the above four constructs and control plasmid. Each chicken was intramuscally immunized with 200 microg plasmid DNA three times in a two-week interval. Two weeks after the third immunization, chickens were injected with H5N1 subtype avian influenza virus. Before the virus loading no detectable antibodies to HA were found in the chicken serum; but high levels of HI antibodies were detected in the serum of the survived chickens. The percentages of CD4+ and CD8+ T lymphocyte in peripheral blood of immunized chickens increased steadily after the vaccination. After virus loading all chickens in the control group died within three to eight days, and the survival rates of the four DNA vaccine groups were as follows: pIRES/HA, 54.5%; pIRES/tHA, 30%, pIRES/ tHA-NPep, 36.3%, pIRES/tHA-NPep-IFN-gamma, 50%. These results indicated that multi-epitopes DNA immunization can induce immune response and protect chickens from homologous virus loading.
Animals
;
Chickens
;
Epitopes
;
immunology
;
Influenza A Virus, H5N1 Subtype
;
immunology
;
pathogenicity
;
Influenza in Birds
;
immunology
;
prevention & control
;
virology
;
Vaccines, DNA
;
immunology
6.Association between Fecal Bile Acids and Colorectal Cancer: A Meta-analysis of Observational Studies.
Jin Lu TONG ; Zhi Hua RAN ; Jun SHEN ; Guo Quan FAN ; Shu Dong XIAO
Yonsei Medical Journal 2008;49(5):792-803
PURPOSE: To provide a systematic review with meta-analysis for addressing the relationship between fecal bile acids (FBAs) and colorectal cancer. MATERIALS AND METHODS: Electronic databases were searched for all observational studies that examined the relationship between FBAs and colorectal cancer or adenoma, and calculated weighted mean difference (WMD) and 95% confidence interval (CI). Publication bias was assessed with funnel plot. RESULTS: Twenty case-control or cohort studies were identified. All studies were pooled to assess the relationship between total FBAs and cancer/adenoma of the large bowel, however, no association was seen (WMD 0.61mg/g freeze-dried feces; 95% CI: -0.35-1.57). Significantly increased concentration of chenodeoxycholic acid (CDCA) was seen while pooling to assess the relationship between CDCA and cancer/adenoma of the large bowel (WMD 0.13 mg/g freeze-dried feces; 95% CI: 0.01-0.25), especially for colorectal cancer (WMD 0.28mg/g freeze-dried feces; 95% CI: 0.10-0.46). However, no significant differences in deoxycholic acid (DCA), lithocholic acid (LCA), and primary and secondary bile acids, were seen between patients with cancer and patients with matched controls regardless of fixed and random effects models. CONCLUSION: CDCA might play a role in the etiology of colorectal cancer.
Bile Acids and Salts/*metabolism
;
Carcinoma/etiology/*metabolism
;
Case-Control Studies
;
Cohort Studies
;
Colorectal Neoplasms/etiology/*metabolism
;
Feces/*chemistry
;
Female
;
Humans
;
Male
7.Role of caspase-8 and DR5 in TRAIL-induced apoptosis of neuroblastoma cells.
Hai-Xia TONG ; Ji-Hong ZHANG ; Li MA ; Chun-Wei LU ; Jin-Hua ZHANG
Chinese Journal of Contemporary Pediatrics 2006;8(4):327-330
OBJECTIVETumor necrosis factor related apoptosis inducing ligand (TRAIL) induces cell death in a variety of tumors but not in normal cells. TRAILdouble ended arrow-resistance of most neuroblastoma (NB) cell lines is related to the loss of caspase-8 expression and the expression and distribution of membrane TRAIL-receptors. This study investigated the role of caspase-8 and DR5 in TRAIL-induced apoptosis of NB cell line SKNDZ.
METHODSThe expression of caspase-8 mRNA was detected by RT-PCR. The expression of DR5 protein was detected by Western Blot analysis. The effects of TRAIL, IFNgamma +TRAIL, chemotherapeutic agent (adriamycin or etoposide) + TRAIL, and chemotherapeutic agent +TRAIL+ IFNgamma on the growth and apoptosis of SKNDZ cells were detected by MTT assay and flow cytometry.
RESULTScaspase-8 was not expressed in SKNDZ cells but IFNgamma treatment resulted in an increase of caspase-8 expression. Expression of DR5 protein was not detected in SKNDZ cells but an increased DR5 protein expression was found after treatment with adriamycin or etoposide. The SKNDZ cells expressing caspase-8 were not sensitive to TRAIL but those SKNDZ cells expressing both caspase-8 and DR5 were sensitive. The early apoptosis rates of the adriamycin /etoposide + IFNgamma+TRAIL groups [(17.9 +/- 3.6)%, (14.8 +/- 3.3)%] were higher than that of the IFNgamma+TRAIL group [(3.9 +/- 1.2)% ](F=26.233, P < 0.01).
CONCLUSIONSSKNDZ cells expressing both caspase-8 and DR5 restored the TRAIL sensitivity. Caspase-8 and DR5 play a key role in TRAIL-induced apoptosis of NB cells.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Blotting, Western ; Caspase 8 ; Caspases ; physiology ; Humans ; Interferon-gamma ; pharmacology ; Membrane Glycoproteins ; pharmacology ; Neuroblastoma ; pathology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Receptors, Tumor Necrosis Factor ; physiology ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; pharmacology ; Tumor Suppressor Protein p53 ; physiology
8.p16 and MGMT gene methylation in sputum cells of uranium workers.
Shi-biao SU ; Lu-jing YANG ; Wei ZHANG ; Ya-li JIN ; Ji-hua NIE ; Jian TONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(2):92-95
OBJECTIVETo study the methylation of O-6-methylguanine-DNA methyltransferase (MGMT) and p16 gene in the sputum cells of radon-exposed population. To provide the experimental base for finding the molecular biomarker of the high risk population of the radon-induced lung cancer.
METHODS91 radon-exposed workers were divided into 4 groups, high dosage group (> 120 WLM), middle dosage group (between 60 and 120 WLM), low dosage group (between 30 and 60 WLB) and lower dosage group (between 2 and 30 WLM) according to the accumulated exposure dosage of the radon daughters. The abnormal methylation of p16 and MGMT gene in the sputum cells of the population in the four groups was detected with the methylation specific PCR (MSP).
RESULTSThere was significantly upward trend for the p16 gene methylation rate (0.00%-20.00%), the MGMT gene methylation rate (0.00%-28.00%) and the total methylation rate (0.00%-40.00%) with the increase of the accumulated exposure dosage of the radon daughters (P < 0.01).
CONCLUSIONThe methylation of p16 and MGMT gene is related to the accumulate exposure dosage of the radon daughters.
Carcinogens, Environmental ; adverse effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; Dose-Response Relationship, Radiation ; Humans ; Male ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; Occupational Exposure ; Radon ; adverse effects ; Radon Daughters ; adverse effects ; Sputum ; metabolism
9.V89L polymorphism of the testosterone 5-alpha-reductase II gene and prognostic factors of prostate cancer.
Ming TONG ; Yan-Yang JIN ; Gang LI ; Si-Ming LIU ; Chun-Dong JI
National Journal of Andrology 2010;16(11):990-993
OBJECTIVETo investigate the association of V89L polymorphism of the SRD5A2 gene with the prognostic factors of prostate cancer (PCa).
METHODSWe identified the V89L polymorphic sites of the SRD5A2 gene after Rsa-1 restriction enzyme digestion, observed the distribution of V89L (VV, VL and LL) polymorphism in 112 PCa and 89 benign prostate hyperplasia (BPH) patients, and determined the association of V89L polymorphism with the age, free PSA (fPSA), total PSA (tPSA), fPSA/tPSA ratio, tumor stage and Gleason score of the PCa patients.
RESULTSNo statistically significant differences were found in the V89L polymorphism-induced genetic risk frequencies between the PCa and BPH groups (chi2 = 3. 606, df = 2, P = 0. 165), nor any significant correlation between the genotypes of VV and VL + LL and the differences in the fPSA, tPSA, fPSA/tPSA ratio, tumor stage, Gleason score and age of the PCa patients. VV and VL + LL showed no obvious association with the prognostic factors of PCa.
CONCLUSIONV89L polymorphism is not related with the prognosis of PCa, but may be indirectly associated with its risk.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase ; genetics ; Aged ; Aged, 80 and over ; Genotype ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Neoplasm Staging ; Polymorphism, Genetic ; Prognosis ; Prostatic Neoplasms ; diagnosis ; genetics ; pathology
10.Combination of gamma-interferon with TRAIL and cisplatin or etoposide induces apoptosis in human neuroblastoma cell line SH-SY5Y.
Hai-Xia TONG ; Chun-Wei LU ; Ji-Hong ZHANG ; Li MA ; Jin-Hua ZHANG
Chinese Medical Sciences Journal 2007;22(1):38-43
OBJECTIVETo study the effect of gamma-interferon (IFNgamma), tumor necrosis factor related apoptosis inducing ligand (TRAIL), and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms.
METHODSThe expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFN-gamma, TRAIL, IFNgamma + TRAIL, IFN-gamma + Caspase 8 inhibitor + TRAIL, IFNgamma + cisplatin + TRAIL, and IFNgamma + etoposide + TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay.
RESULTSCaspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNgamma. SH-SY5Y cells themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNgamma were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFN-gamma + TRAIL group was significantly higher than those of control group, IFN-gamma group, TRAIL group, and inhibitor group (P < 0.01). There was no significant difference among IFN-gamma + TRAIL group, IFNgamma + cisplatin + TRAIL group, and IFNgamma + etoposide + TRAIL group.
CONCLUSIONSIFNgamma could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 8 ; genetics ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Etoposide ; pharmacology ; Humans ; Interferon-gamma ; pharmacology ; Neuroblastoma ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology