2.Relationship between the expression of Cofilin in alveolar and peripheral blood mononuclear cells in COPD patientsand the function of phagocytic aspergillus
Xiping WU ; Ze ZHU ; Zekui FANG ; Xin CHEN
The Journal of Practical Medicine 2017;33(13):2129-2132
Objective To investigate the expression of Cofilin in alveolar and peripheral blood mononuclear cells in COPD patients and the correlation with the function of phagocytic aspergillus. Methods 20 COPD pa-tients were selected from July 2015 to May 2016 in the outpatient department of respiratory medicine of Zhujiang Hospital of Southern Medical University ,then divided into the experimental group 1(group Aand B in 2011 version of GOLD),group 2(test group C and group D in 2011 version of GOLD),and the healthy control group in 10 people. The AM and MDM in the peripheral bloodwere extracted respectively in the 3 groups by bronchoalveolar lavage and,and the ability of AM and MDM in each group were detected. The expression of Cofilin protein was measured by real-time fluorescence quantitative(qRT-PCR) and Western blotting ,and the differenceswere compared among the three groups. Results The colony numbers of MDM/AM in the 3 groups were 17 ± 3,16 ± 2, 42 ± 3(F = 73.446 ,P < 0.001),and the colony numbers of MDM/AM in the two test groupshad significantly different from the healthy group ,while no significant difference was found in the two test groups. The results of fluorescence quantitative reverse transcription polymerase chain reaction and Western blot analysis showed that the expression of Cofilin in the two test groups was significantly higher than that in the healthy group. Conclusion The decreased function of phagocytic aspergillus in the alveolar and peripheral blood mononuclear cells in COPD patients may relates to the increase of cofilin.
3.Development of a novel screening assay for inhibitors targeting HIF-1alpha and P300 interaction.
Fang-Fang LAI ; Fei NIU ; Han-Ze YANG ; Wan-Qi ZHOU ; Xiao-Guang CHEN
Acta Pharmaceutica Sinica 2014;49(6):849-853
Hypoxia is a general characteristic of most solid malignancies and intimately related to cancer progression. Homeostatic response to hypoxia is primarily mediated by hypoxia inducible factor-1alpha (HIF-1alpha) that elicits transcriptional activity through recruitment P300 coactivator. Targeting the interaction of HIF- alpha and P300 would thus constitute a novel approach for cancer treatment by suppressing tumor angiogenesis and metastasis. Here, a screening assay was developed for inhibitors targeting the interaction between HIF-1alpha and P300. The nucleotide sequence of human HIF-1alpha and P300 were cloned into pBIND and pACT vectors, named pBIND-HIF1alpha and pACT-P300. The interaction of HIF-1alpha and P300 was identified in HEK293 cell using mammalian two-hybrid system. And compound chetomin decreased their interaction in this mammalian two-hybrid system. We further verified HIF-1 inhibition effect of chetomin in U251-HRE cells. Therefore, we established a screening assay combined HIF-1alpha and P300 mammalian two-hybrid system and U251-HRE reporter assay for HIF-1 selective inhibitors.
Cell Hypoxia
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Disulfides
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pharmacology
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Drug Screening Assays, Antitumor
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E1A-Associated p300 Protein
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antagonists & inhibitors
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HEK293 Cells
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit
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antagonists & inhibitors
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Indole Alkaloids
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pharmacology
;
Two-Hybrid System Techniques
4.Development of infusion tele-control system based on LonWorks
Zifeng CHENG ; Duanyun PENG ; Ze WEI ; Xin WANG ; Shaolin MA ; Hongbo CHEN ; Ting FANG
Chinese Medical Equipment Journal 2004;0(08):-
This paper introduces some information of the infusion tele-control system based on LonWorks, including its system function, operation principle, software and hardware designs. With high reliability, low cost and convenient operation, this system has a bright future in the market and its application.
5.Cloning and Overexpression of Phytase Gene appA form Escherichia coli
Yin CHEN ; Zhong-Ze ZHU ; Zhi-Fang ZHANG ; Jia-Lu HE ;
Microbiology 1992;0(03):-
A strain Escherichia coli with high production of phytase was screened from pig excreta. Phytase gene appA, with 1,299 bp coding region in full length, was cloned from its genome by polymerase chain reaction (PCR) . The gene appA was then cloned into the prokaryotic expression vector pET-28a ( + ) . In the host BL21, the phytase appA was overexpressed by shaker-cultivation (up to 692 U/mL) . The enzymatic analysis of the prokaryotic derived appA phytase revealed that its optimal pH and temperature was 4.5 and 60℃, respectively.
6.Research on differentially expressed genes related to substance and energy metabolism between healthy volunteers and splenasthenic syndrome patients with chronic superficial gastritis.
Yang ZE-MIN ; Chen WEI-WEN ; Wang YING-FANG
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(2):159-163
OBJECTIVETo analyze the metabolic states of the lipids, protein, carbohydrate, and nucleic acid for chronic superficial gastritis patients of splenasthenic syndrome (SS), and to explore the pathogenesis mechanism of SS based on substance and energy metabolisms.
METHODSDuring June 2004 to March 2005, recruited were four chronic superficial gastritis patients of SS who visited at the First Hospital of Guangzhou University of Chinese Medicine and Guangdong Provincial Hospital of Traditional Chinese Medicine. Four healthy volunteers were recruited from Guangzhou University of Chinese Medicine. Their gastric mucosa was extracted to perform experiments of DNA microarray. The dual-channel DNA microarray data were mined and bioinformatics analyzed by BRB ArrayTools and IPA software.
RESULTSFifteen genes were involved in substance and energy metabolisms in 20 differentially expressed genes, accounting for 75%.Among these genes, one gene was up-regulated, 14 genes down-regulated, and 11 genes were enzyme gene. Differentially expressed genes related to lipid metabolism included ACAA2 and CYP20A1, manifested as fatty acid catabolism and cholesterol transformation. Genes related to protein metabolism included ALDH9A1, ASL, ASS1, PCY-OX1L, RPS28, UBE2D2, UBXN1, B3GNT1, GCNT1, and PPP1R3C, manifested as decreased amino acid metabolism that may affect the biologic processes such as autonomic nerve, urea cycle, etc., reduced protein synthesis, increased ubiquitination of fault fold proteins, and decreased post-translated modification of glycosylation and dephosphorylation. Genes related to carbohydrate metabolism included PPP1R3C, B3GNT1, and GCNT1, manifested as decreased glycogen and glycan syntheses. Genes related to nucleic acid metabolism included RMI1, SMARCD3, and PARP1, manifested as degraded DNA duplication and transcription, and increased DNA damage repair.
CONCLUSIONSThe metabolisms of the lipids, protein, carbohydrate, and nucleic acid in chronic superficial gastritis patients of SS obviously decreased, manifested mainly as down-regulated enzyme gene expression. We inferred that these might be one of the vital pathogenesis mechanisms for nutrition dysmetabolism of SS.
Adult ; Case-Control Studies ; Energy Metabolism ; genetics ; Female ; Gastritis ; diagnosis ; genetics ; metabolism ; Gene Expression ; Gene Expression Profiling ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Oligonucleotide Array Sequence Analysis ; Young Adult
7.Study on differential estrogen receptor beta expression of mandibular condylar chondrocyte in temporomandibular joint osteoarthritis.
West China Journal of Stomatology 2006;24(5):469-472
OBJECTIVETo test the differential Estrogen Receptor (ER) beta levels of mitochondrial proteome of mandibular condylar chondrocyte in the rat model of temporomandibular joints osteoarthritis (TMJOA).
METHODS25 SD rats were divided into experimental group (15 rats) and control group (10 rats) randomly. TMJOA models were created in left sides of TMJ of 15 SD rats by the partial resection of the articular disc. The experimental rats were killed 3 months after operation. After the chondrocytes culture, Immunohistochemistry, semiquantitative RT-PCR and Western blot were used to test the differential ER beta expression levels in mitochondrial proteome of mandibular condylar chondrocytes. Mitochondrial proteins identification was carried out by two-dimensional electrophoresis and peptide mass fingerprint (PMF).
RESULTSPMF showed that one of the differential proteins was ERP. Immunohistochemistry and Western blot results suggested the significant difference of ERbeta protein levels between operation-treated and control group. The operation-treated group had lower ERbeta levels (P < 0.01).
CONCLUSIONIt has been demonstrated that ERbeta protein levels were decreased in mitochondria of TMJOA mandibular condylar chondrocytes, which suggests a role for mitochondrial ERbeta in the effects on TMJOA. The pathological role of ERbeta in the regulation of TMJOA progress remains to be elucidated.
Animals ; Cartilage, Articular ; Chondrocytes ; Estrogen Receptor beta ; Immunohistochemistry ; Mandibular Condyle ; Osteoarthritis ; Rats ; Rats, Sprague-Dawley ; Temporomandibular Joint
8.Study on gene differential expressions of substance and energy metabolism in chronic superficial gastritis patients of Pi deficiency syndrome and of pi-wei hygropyrexia syndrome.
Ze-Min YANG ; Wei-Wen CHEN ; Ying-Fang WANG
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(9):1180-1187
OBJECTIVETo analyze the metabolic levels of energy and substance in chronic superficial gastritis (CSG) patients of Pi deficiency syndrome (PDS) and of Pi-Wei hygropyrexia syndrome (PWHS), including lipid, protein, nucleic acid, carbohydrate, trace element, and energy metabolism, and to study the pathogenesis mechanism of PDS from substance and energy metabolisms.
METHODSRecruited were 8 CSG patients who visited at First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine and Guangdong Provincial Hospital of Traditional Chinese Medicine from June 2004 to March 2005, including 4 patients of PDS and 4 of PWHS. Their gastric mucosae were used for experiments of DNA microarray. The dual-channel DNA microarray data were bioinformatically analyzed by BRB ArrayTools and IPA Software.
RESULTSObtained were fifty-six differentially expressed genes involved in substance and energy metabolisms with the expression fold more than 2, including 11 genes up-regulated and 45 genes down-regulated. Of them, genes correlated to lipid metabolism included CRLS1, LRP11, FUT9, GPCPD1, PIGL, SULT1A4, B3GNT1, ST8SIA4, and ACADVL, mainly involved in the metabolic processes of fatty acid, cholesterol, phospholipids, and glycolipid. Genes correlated to protein metabolism included ASRGL1, AARSD1, EBNA1BP2, PUM2, MRPL52, C120RF65, PSMB8, PSME2, UBA7, RNF11, FBXO44, ZFYVE26, CHMP2A, SSR4, SNX4, RAB3B, RABL2A, GOLGA2, KDELR1, PHPT1, ACPP, PTPRF, CRKL, HDAC7, ADPRHL2, B3GNT1, ST8SIA4, DDOST, and FUT9, mainly involved in the biosynthesis processes of protein, ubiquitination, targeted transport and post-translation modification. Genes correlated to nucleic acid metabolism included DFFB, FLJ35220, TOP2A, SF3A3, CREB3, CRTC2, NR1D2, MED6, GTF2IRD1, C1ORF83, ZNF773, and ZMYND11, mainly involved in DNA replication and repair, transcription regulation. Genes correlated to carbohydrate metabolism included AGL, B3GNT1, FUT9, ST8SIA4, SULT1A4, DDOST, and PIGL, mainly involved in glucogen degradation and glycoconjugate biosynthesis. Genes correlated to trace element metabolism included COMMD1, SLC39A6, FTL, CHRFAM7A, SCGN, and S100A6, mainly involved in ion metabolisms of copper, zinc, ferri, and calcium. Genes correlated to energy metabolism included AK3 and COX7B, mainly involved in mitochondria structure and oxidative phosphorylation processes.
CONCLUSIONThe metabolic levels of energy and substance including lipid, protein, nucleic acid, carbohydrate, and trace element were obviously reduced in patients of PDS, which might be an important pathogenesis mechanism for its occurrence.
Adult ; Energy Metabolism ; genetics ; Female ; Gastritis ; diagnosis ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Transcriptome
9.Expressions of homing-related adhesion molecules in hematopoietic stem/progenitor cells derived from human placenta, umbilical cord arterial and venous blood.
Rui SU ; Dai-Xiong CHEN ; Ning FANG ; Qi CHEN ; Fang-Ze GONG
Journal of Experimental Hematology 2006;14(3):582-586
The aim of this study was to evaluate the homing capabilities of hematopoietic stem/progenitor cells (HSPCs) derived from human placenta tissues (PT). Single cell suspension of human PT was prepared by mechanical method. The expression levels of homing-related adhesion molecules (HRAM) including CD11a, CD49d, CD44, CD49e, CD62L and CD54 on CD34(+) cells and the percentages of CD34(+) cells and their subpopulations in nucleated cells (NC) from fresh human PT, umbilical cord arterial blood (UCAB) and umbilical cord venous blood (UCVB) were detected by using flow cytometry. The results showed that the percentage of CD34(+) cells and CD34(+)CD38(-) cells in placenta were higher than those in UCAB and UCVB. There were no significant difference in percentage of HSPC between UCAB and UCVB. Placenta-derived CD34(+) cells strongly expressed CD11a, CD49d, CD44, CD49e and CD54, among which expression levels of CD49e and CD54 on placenta-derived CD34(+) cells were significantly higher than those on UCAB and UCVB-derived CD34(+) cells. While the percentage of CD34(+)CD62L(+) cells in placenta was only lower than that in UCVB. It is concluded that human placenta is rich in HSPC. Moreover, the expression levels of most HRAM in CD34(+) cells from PT are higher than those from UCAB and UCVB or are close to them. It suggested that HSPCs derived from PT might have stronger homing capabilities than those from UCB.
Antigens, CD34
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biosynthesis
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Cell Adhesion Molecules
;
biosynthesis
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Fetal Blood
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cytology
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Hematopoietic Stem Cells
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metabolism
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Humans
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Hyaluronan Receptors
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biosynthesis
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Integrin alpha5
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biosynthesis
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Intercellular Adhesion Molecule-1
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biosynthesis
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Placenta
;
cytology
10.Efficacy analysis of sacral canal injection in patients with lumbar disc herniation associated with non-sciatica.
Jian CHEN ; Gan-Jun WEN ; Lin-Fang ZENG ; Pei-Ru XIAO ; Ze-Qun CHEN ; Yikai LI
China Journal of Orthopaedics and Traumatology 2013;26(8):668-671
OBJECTIVETo observe the outcome after sacral canal injection in patients with disc herniation associated with without sciatica.
METHODSFrom December 2010 to June 2011, 65 patients with acute low back pain without sciatica due to lumbar disc herniation or bulging confirmed by CT or MRI were randomly divided into sacral canal injection group (experimental group) and lumbar oblique wrench group (control group): the experimental group had 35 cases, including 30 males and 5 females, with an average age of (43.90 +/- 1.14) years old ranging from 33 to 56 years old. The control group had 30 cases, including 27 males and 3 females,with an average age of (44.00 +/- 1.19) years old ranging from 34 to 57 years old. The course of morbidity was 1 to 3 days. All patients received sacral canal injection or lumbar oblique wrench method. The visual analog scale (VAS) scores before and at 30 min after treatment were compared between two groups.
RESULTSThe symptom of acute low back pain were relieved obviously. The average VAS scores before and after treatment in experimental group were decreased from 6.63 +/- 0.97 to 3.06 +/- 1.51,in control group were from 6.67 +/- 0.96 to 3.93 +/- 1.20 respectively. These two methods could improve the VAS score,but the effect of sacral canal injection group was better than that of lumbar oblique wrench group, there was statistically differences (P < 0.05).
CONCLUSIONIt is effective that the methods of sacral canal injection and lumbar oblique wrench applied to patients with acute low back pain without sciatica due to lumbar disc herniation or bulging confirmed, the former has better effect.
Adult ; Female ; Humans ; Injections, Spinal ; Intervertebral Disc Displacement ; drug therapy ; Lidocaine ; administration & dosage ; Lumbar Vertebrae ; Male ; Middle Aged ; Prednisone ; administration & dosage ; Sacrococcygeal Region ; Visual Analog Scale