1. Expression and clinical significance of hypoxia-inducible factor 1a and vascular endothelial growth factor in endometrial cancer
Academic Journal of Second Military Medical University 2019;40(4):459-463
Objective To investigate the difference of hypoxia-inducible factor 1a (HIF-1a) and vascular endothelial growth factor (VEGF) expression in endometrial cancer and para-carcinoma tissues, and to explore the clinical significance of hypoxia and the two proteins in the development and progression of endometrial cancer. Methods From Jan. 2011 to Dec. 2012, 128 patients with endometrial cancer underwent surgery in Tongji Hospital of Tongji University. The expression of HIF-1a and VEGF in cancer tissues and paired para-carcinoma tissues was detected using immunohistochemical method. The patients were followed up regularly, and the relationship between the expression of HIF-1a and VEGF and the prognosis of the patients was analyzed. The hypoxic cell model of human endometrial cancer was constructed to detect the expression of HIF-1a and VEGF proteins and observe the cell proliferation, invasion and apoptosis. Results The positive rates of HIF-1a and VEGF in cancer tissues were significantly higher than those in the para-carcinoma tissues (both P<0.05). The positive rate of HIF-1a was higher in the patients with lymph node metastasis, high histological grade, maximal tumor dimeter=4 cm or positive progesterone receptor (all P<0.05). The positive rate of VEGF was higher in the patients with lymph node metastasis, high histological grade, deep myometrial invasion, maximal tumor dimeter=4 cm, positive estrogen receptor, positive progesterone receptor or high pathological stage (all P<0.05). The 5-year overall survival rate of the patients with negative HIF-1a was significantly higher than that of the patients with positive HIF-1a (P<0.05), while there was no significant difference in the 5-year overall survival rate between the patients with negative and positive VEGF (P>0.05). In the hypoxic cell model of human endometrial cancer, the expression levels of HIF-1a and VEGF were significantly increased, cell proliferation and invasion were significantly increased, and the cell apoptosis was significantly reduced (all P<0.05). Conclusion HIF-1a and VEGF are related to the progress of endometrial cancer, and positive expression of HIF-1a indicates a poor prognosis.
2.Effect of selected fungi on the reduction of gossypol levels and nutritional value during solid substrate fermentation of cottonseed meal.
Wen-ju ZHANG ; Zi-rong XU ; Jian-yi SUN ; Xia YANG
Journal of Zhejiang University. Science. B 2006;7(9):690-695
The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fermentation of cottonseed meal (CSM). Six groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. capsuligena ZD-1, C. tropicalis ZD-3, S. cerevisae ZD-5, A. terricola ZD-6, A. oryzae ZD-7, or A. niger ZD-8. One not inoculated group (substrate) was used as a control. Levels of initial and final free gossypol (FG), crude protein (CP), amino acids (AA) and in vitro digestibility were assayed. The experiment was done in triplicate. The experimental results indicated that microbial fermentation could greatly decrease (P<0.05) FG levels in CSM. The detoxification efficiency differed between the species of microorganisms applied. From the perspective of reducing CSM potential toxicity, C. tropicalis ZD-3 was most successful followed by S. cerevisae ZD-5 and A. niger ZD8. They could reduce FG levels of CSM to 29.8, 63.07 and 81.50 mg/kg based on DM (dry matter), respectively, and their detoxification rate were 94.57%, 88.51% and 85.16%, respectively. If crude protein, amino acids content and their in vitro digestibility were also taken into account, A. niger ZD-8 may be the best choice. The CP content of CSM substrate fermented by C. tropicalis ZD-3 and A. niger ZD-8 were improved by 10.76% and 22.24%; the TAA (total amino acids) contents were increased by 7.06% and 11.46%, and the EAA (essential amino acids) were raised by 7.77% and 12.64%, respectively. Especially, the levels of methionine, lysine and threonine were improved greatly (P<0.05). The in vitro CP digestibility of CSM fermented by C. tropicalis ZD-3 and A. niger ZD-8 was improved by 13.42% and 18.22%, the TAA were increased by 17.75% and 22.88%, and the EAA by 16.61% and 21.01%, respectively. In addition, the in vitro digestibility of methionine, lysine and threonine was also improved greatly (P<0.05).
Amino Acids
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analysis
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Cottonseed Oil
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chemistry
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metabolism
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Digestion
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Fermentation
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Fungi
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metabolism
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Gossypol
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analysis
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Nutritive Value
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Plant Proteins
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analysis
3.Gloning and Sequence Analysis of Dienelactone Hydrolase Gene
Wen-Hui ZHONG ; Ming SUN ; Guo-Qing HE ; Xiao-Shan FENG ; Zi-Niu YU ;
Microbiology 1992;0(03):-
A 2,4 -dichlorophenol degrading Pseudomonas strain GI241-1 was isolated from a soil sample. The dienelactone hydrolase gene, designated as dcpD which encodes dienelactone hydrolase involved in transforming cis-2-chloro-dienelactone into 2-chloromaleylacetic acid, was cloned from this bacterium strain. The gene cloning strategy was to construct genomic library after location of its neighbouring gene by Southem blot and to screen the aim transformant by dot blotting. Sequencing results showed that length of dcpD is 702bp. The sequence of dcpD and the deduced amino acid are different from the relative sequences registered in the GenBank.
4.Expression of human mitochondrial transcription termination factor-3 in non-small-cell lung cancer and its clinicopathological significance
Jiaji ZI ; Yongqin YANG ; Meitao SUN ; Wen MEI ; Xiaojuan ZHANG ; Wei XIONG
Journal of Medical Postgraduates 2017;30(2):160-164
Objective The purpose of this study was to investigate the expression of human mitochondrial transcription termi-nation factor-3 ( hMTERF3) in non-small cell lung cancer ( NSCLS) and to analyze its clinicopathological significance. Methods The paraffin block samples used in this study included 65 cases of NSCLC and 32 cases of normal alveolar epithelial tissues. We determined the expressions of hMTERF3 in NSCLC and normal alveolar epithelial tis-sues by immunohistochemistry, calculate the survival rate using the Kaplan-Meier method, and analyzed the risk factors affecting the prognosis of NSCLC using the Cox Proportional Hazard Model. Results In the 65 cases of NSCLC, 31 ( 47. 69%) showed positive expression of hMTERF3. The total survival time was significantly shor-ter in the patients with a high than in those with a low hMTERF3 ex-pression ([30.39±3.35] vs [57.61±7.12] mo, P<0.05). The riskfactors affecting the prognosis of NSCLC included positive expression of hMTERF3 (HR=3.302, 95% CI:1.598-6.905) and lymph node metastasis (HR=4.052, 95% CI: 1.212-12.398). Conclusion hMTERF3 is overexpressed in NSCLC. Highly expressed hMTERF3 and lymph node metastasis reduce the survival time of NSCLC patients, suggesting that hMTERF3 may be a potential bio-marker for the prognosis of NSCLC.
5.Finite element analysis of intertrochanteric fractures in older adults based on Hypermesh 14.0 and LS-DYNA software
Xiang-Xin HE ; Zi-Ling LIN ; Peng-Fei LI ; Gen-Fa DU ; Wen-Tao SUN ; Xin-Min CHEN ; Zi-Yi LIANG
Chinese Journal of Tissue Engineering Research 2018;22(11):1725-1730
BACKGROUND: At present, finite element analysis can be used to judge intertrochanteric fractures, but mostly limited in the distribution of stress. Finite element model of various intertrochanteric fractures has not been reported in detail.OBJECTIVE: To build various types of intertrochanteric fracture models with Hypermesh 14.0 and LS-DYNA software to simulate the falling-induced external force on proximal femur, and to evaluate the effect of models, and to analyze the biomechanical mechanism of intertrochanteric fractures. METHODS: Normal side CT image data of one case of elderly intertrochanteric fracture were collected and imported into Mimics software to establish the proximal femur geometric models, were then analyzed and operated by LZ-DYNA solver after imported into Geomagic studio 2013 and Hypermesh 14.0 for smoothing and meshing. Before analysis, the material parameters were set, the boundary conditions were confirmed, and given the loading parameters. The operating results were checked in Hyper View. RESULTS AND CONCLUSION: (1) The distribution of stress of proximal femur exactly matched to the previous study. EvansⅠtype intertrochanteric fracture model was obtained under continuous shear stresses, and six types of fractures were obtained by adjusting the load. (2) These results manifest that based on the Hypermesh 14.0 and LS-DYNA software, the finite element can well simulate the intertrochanteric fractures, and shear stress plays an important role in intertrochanteric fractures, which can provide experimental basis for the prevention and treatment of intertrochanteric fractures.
6.Congenital cystic adenomatoid malformation of fetus:manifestations and diagnosis of MRI
Zi-Yan SUN ; Li-Ming XIA ; Xin-Lin CHEN ; Cheng-Yuan WANG ; Xiao-Hong YANG ; Wen-Zhong YANG ;
Chinese Journal of Radiology 2001;0(05):-
Objective To demonstrate the MRI manifestations of congenital cystic adenomatoid malformation(CCAM)and to evaluate the diagnostic value of MRI.Methods Thoracic axial,sagittal and coronal plane scanning were performed with SSFSE in 9 fetuses diagnosed or suspected of CCAM by ultrasound(US)within 1—2 days after US examination.The diagnosis was confirmed by postnatal autopsy or follow-up.Results In nine fetuses,seven cases of CCAM were diagnosed with MRI and confirmed by autopsy,one case was congenital pulmonary sequestration,one was normal on MRI and two weeks late US.In seven cases of CCAM(5 males and 2 females),four cases were in the right side,three in the left.Four cases of macrocystic type CCAM showed single or multiple cystic long T_2 signal in the unilateral lung,three cases of microcystic type CCAM presented long T_2 signal without cyst.Vessels with flow void phenomenon were found in 2 cases of CCAM and 1 case of pulmonary sequestration.Conclusion MRI can clearly show the location,range and contour of CCAM.The vessel originated from the aorta is suggestive of pulmonary sequestration.
7.Targeted imaging ability of a biotinylated imaging probe Biotin-S-S-Rhodol for breast cancer cells in vitro.
Bi-Juan WU ; Xing-Zi ZHOU ; Jing-Wen SUN ; Cui-Wen TAN ; Xin-Rong WU
Journal of Southern Medical University 2017;37(1):124-129
OBJECTIVETo investigate performance of a biotinylated imaging probe 3a for targeted imaging of breast cancer cells.
METHODSUltraviolet absorption spectrum and fluorescence spectrum were employed to analyze the spectral characteristics of 3a. The fluorescence spectrums of 3a treated with different concentrations of glutathione (GSH) were obtained to determine the sensibility of 3a to GSH. Flow cytometry was used to determine the cellular uptake of 3a by MCF-7 cells, MDA-MB-231 cells and Hs 578Bst cells in the presence or absence of biotin, and the imaging performance of 3a in the 3 cell lines was assessed under an inverted fluorescent microscope. The toxicity of 3a to the cells was evaluated using MTT method.
RESULTS3a showed the strongest absorption peak at 510 nm, and its fluorescence emission signal was the strongest at 544 nm. As the concentration of GSH increased (0-6 mmol/L), 3a exhibited an increasing fluorescence signal at 544 nm. The cellular uptake of 3a was markedly higher in MDA-MB-231 cells and MCF-7 cells than in Hs 578Bst cells. The imaging studies showed that 3a had a good breast cancer cell-targeting property and produced clear images under fluorescent microscope. MTT assay demonstrated no obvious toxicity of 3a in Hs 578Bst cells even at the concentration of 20 µmol/L, but MCF-7 cells and MDA-MB-231 cells exposed to 2-20 µmol/L 3a showed a lowered cell viability.
CONCLUSION3a is capable of targeted imaging of breast cancer cells mediated by biotin. 3a at the concentration of 2-20 µmol/L has minimal cytotoxicity to normal breast cells but can lower the viability of breast cancer cells.
9.Influence of high mobility group box 1 on migration of human cord blood CD34(+) cells.
Xin CHEN ; Xing-Bing WANG ; Hui-Lan LIU ; Wen YAO ; Kai-Di SONG ; Zi-Mi SUN
Journal of Experimental Hematology 2009;17(2):422-425
The objective of study was to explore the influence of high mobility group box 1 (HMGB1) on migration of cord blood CD34(+) cells and their mechanism of migration. The expressions of receptor for advanced glycation end products (RAGE), toll-like receptor-2 (TLR2) and TLR4 were detected by flow cytometry. The CD34(+) cells in umbilical cord blood (CB) were enriched by MiniMACS and were exposed to various concentration of HMGB1 (10, 50, 100, 1, 000 ng/ml), then the migration effect of HMGB1 on umbilical cord blood (UCB) CD34(+) cell count was determined by microscopy, the chemotactic index was calculated. The CD34(+) cells untreated with HMGB1 were used as control. The results indicated that the purity of the isolated CD34(+) cells was more than 98%. The HMGB1 could promote the migration of CD34(+) cells, and the migration effect of HMGB1 on CD34(+) cells in certain concentrations gradually increased along with raise of concentration, the strongest effect was observed in concentration of 100 ng/ml, there was significant difference as compared with control (p < 0.01). Anti-RAGE antibody partially inhibited the migration effect of HMGB1 on CD34(+) cells. It is concluded that the HMGB1 in certain concentration can enhance migration of CD34(+) cells, which may be mediated through RAGE.
Antigens, CD34
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Cell Movement
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drug effects
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Cells, Cultured
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Female
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Fetal Blood
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cytology
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drug effects
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HMGB1 Protein
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pharmacology
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Humans
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Receptor for Advanced Glycation End Products
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Receptors, Immunologic
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metabolism
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Signal Transduction
10.Isolation of 2,4-dichlorophenol degrading bacterium strain and cloning and expression of its 2,4-dichlorophenol hydroxylase gene.
Wen-Hui ZHONG ; Ming SUN ; Guo-Qing HE ; Xiao-Shan FENG ; Zi-Niu YU
Chinese Journal of Biotechnology 2004;20(2):209-214
2,4-Dichlorophenol is toxic and biorefratory organic pollutant. A 2,4-dichlorophenol degrading bacterial strain GT241-1, identified as Pseudomonas sp., was isolated from soil samples which was collected from drainage area of several 2,4-dichlorophenol producing factories. Strain GT241-1 had strong 2,4-dichlorophenol degrading ability, it could decompose 91% 2, 4-dichlorophenol of 90 mg/L within 48 hours at 25 - 30 degrees C, and could utilize 2,4-dichlorophenol, 2,4-dichlorophenoxyacetic acid (2,4-D), benzoate and catechol as sole carbon and energy source. Southern blot showed that 2,4-dichlorophenol hydroxylase gene (dcpA) of strain GT241-1 locates on the about 10kb EcoR I/Xba I fragment. This fragment was recovered, linked to the vecter pUC19 and transformed into the E. coli DH5alpha. A aim transformant, Z539, was obtained by dot blotting from about 1200 transformants. PCR and the sequencing results shew that the whole dcpA gene is contained within the 10kb EcoR I /Xba I fragment of pZ539. This fragment was shortened to about 2.4kb by HindmIII. The shorted fragment was subcloned to vecter pRSET-B to get a transformant BS1-12. The subcloned fragment was sequenced. Sequencing results showed that the whole length of the subcloned fragment containing dcpA is 2389bp and the nucleotide span of coding region is from number 276 to number 2072 (1797 bp), with ATG and TAA as start and stop codon respectively. The sequence analysis of dcpA and the deduced amino acid encoded by dcpA showed that they are different from the relative sequences registered in the GenBank. The subcloned fragment carry the promoter of dcpA, this can deduce from the fact that the upflow length of dcpA coding region is 275bp, and further confirmed by the 2,4-dichlorophenol hydroxylase activity measurement results. The 2,4-dichlorophenol hydroxylase activity of transformant Z539 and BS1-12 were detected, the results showed these transformants have 2,4-dichlorophenol hydroxylase activity. By comparison, the activity of these transformants were lower than that of the strain GT241-1.
Amino Acid Sequence
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Bacterial Proteins
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genetics
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metabolism
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Biodegradation, Environmental
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Chlorophenols
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metabolism
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Cloning, Molecular
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Environmental Pollutants
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metabolism
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Mixed Function Oxygenases
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genetics
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metabolism
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Molecular Sequence Data
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Pseudomonas
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enzymology
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genetics
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isolation & purification
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Soil Microbiology