1.Study of the mechanism of mast cell increase in cellular leiomyoma of uterus
Jie-Qiang LV ; Xue-Qiong ZHU ; Ke DONG ; Mei XIANG ; Yi LIN ; Yue HU ;
Chinese Journal of Obstetrics and Gynecology 2001;0(06):-
Objective To study the mechanism of mast cell increase in cellular leiomyoma of uterus.Methods Tissue sections from 30 cases of cellular leiomyoma of uterus,15 cases of leiomyosarcoma and 30 cases of ordinary leiomyoma were studied using immunohistochemical double labeling techniques.The expression of mast cell tryptase ahd Ki-67 as well as mast cell tryptase and chemotactic factors RANTES,Eotaxin,monocyte chemoattractant protein-1(MCP-1),transforming growth factor-? (TGF-?)were double immunostained.Results Ki-67 in mast cells was rarely expressed in each group. Expressions of regulate upon activation normal T cell expressed and secreted(RANTES),Eotaxin and TGF-? in cellular leiomyoma were 78%,89%,91%,respectively.They were all higher than those in ordinary leiomyoma and leiomyosarcoma(P
2.Effect of ribosome engineering on butenyl-spinosyns synthesis of Saccharopolyspora pogona.
Lin'gen LUO ; Yan YANG ; Hui WEI ; Jie RANG ; Qiong TANG ; Shengbiao HU ; Yunjun SUN ; Ziquan YU ; Xuezhi DING ; Liqiu XIA
Chinese Journal of Biotechnology 2016;32(2):259-263
Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
Genetic Engineering
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Macrolides
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metabolism
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Point Mutation
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Ribosomal Proteins
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genetics
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Ribosomes
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metabolism
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Saccharopolyspora
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metabolism
3.Application of Adjustable Interatrial Fistulization in Operation of Congenital Heart Disease Accompany with Severe Pulmonary Arterial Hypertension
kai-hu, SHI ; xin, CHEN ; hong-wei, SHI ; xin-wei, MU ; li-qiong, XIAO ; hai-peng, ZHAO ; jun-jie, SHAO
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To investigate the method and value of adjustable interatrial fistulization in the operation of congenital heart disease(CHD) accompany with severe pulmonary arterial hypertension(PH).Methods Twenty-seven patients(19 male,8 females) accompany with severe PH were entered the study,age ranged from 4 to 14 years old,weight from 13.7 to 42.0 kilogram.The enrolled diseases included 11 cases of atrial septal defect(ASD),10 cases of ventricular septal defect(VSD),4 cases of patent ductus arteriosus(PDA),and 2 cases of Ebstein syndrome accompany with severe tricuspid insufficiency.All patients were diagnosed as CHD accompany with severe PH(bidirectional shunt)which was the contraindications for routine operation before operation through chest X-ray,electrocardiography,ultrasonic cardiography,cardiac catheteri-zation and cardiac angiography.Results With adjustable interatrial fistulization and treatment to the abnormalities,14 fistulaes were closed immediately after operation,7 fistulaes were closed 2 days after operation,3 fistulaes were closed 3 days and 1 fistulae was closed 4 days after operation and accompanied with empyema discharged initiatively.One fistula was never closed,1 case died from low cardiac output symptom.The effective rate was 92.6%,closed to that of routine operations.Conclusion Adjustable interatrial fistulization is an easy procedure,and it can decrease the danger of PH post-operation effectively and provide operation opportunity for those patients with CHD approaching terminal stage.
4.The Protective Effect of Sodium Hyaluronate on the Cartilage of Rabbit Osteoarthritis by Inhibiting Peroxisome Proliferator-Activated Receptor-Gamma Messenger RNA Expression.
Jian Lin ZHOU ; Shi Qing LIU ; Bo QIU ; Qiong Jie HU ; Jiang Hua MING ; Hao PENG
Yonsei Medical Journal 2009;50(6):832-837
PURPOSE: The purpose of this study was to study the protective effect and influence of sodium hyaluronate (Na-HA) on mRNA expression of peroxisome proliferators-activated receptor gamma (PPAR-gamma) in cartilage of rabbit osteoarthritis (OA) model. MATERIALS AND METHODS: Forty eight white rabbits were randomly divided into A, B, and C groups. Group A was normal control group, B and C groups underwent unilateral anterior cruciate ligament transection (ACLT). The rabbits in group B were injected normal saline after ACLT; and Group C received intraarticular1% sodium hyaluronate (HA) injection 5 weeks after surgery, 0.3 mL once a week. At 11th week after surgery, all the rabbits were sacrificed. The cartilage changes on the medial femoral condyles were graded separately. Cartilage sections were stained with safranin-O and HE, and messenger RNA (mRNA) expression of PPAR-gamma was detected by using real time polymerase chain reaction (Real Time-PCR). RESULTS: Cartilage degeneration in group B was significantly more severe than in A and C injection group. The grey value of Safranin-O of B group was higher than A and C groups. Expression of PPAR-gamma mRNA in group B was higher than group A and C. CONCLUSION: This study shows that Na-HA has a protective effect on articular cartilage degeneration, and the inhibitory effect on the PPAR-gamma mRNA expression may be one of therapeutic mechanism of Na-HA.
Animals
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Cartilage/*drug effects/*metabolism
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Gene Expression/drug effects/genetics
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Hyaluronic Acid/*pharmacology/therapeutic use
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Microscopy
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Osteoarthritis/*drug therapy/metabolism
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PPAR gamma/*genetics
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RNA, Messenger/*genetics
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Rabbits
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Random Allocation
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Reverse Transcriptase Polymerase Chain Reaction
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Viscosupplements/*pharmacology/therapeutic use
5.External ear reconstruction using expanded postauricular scar flap and Medpor framework.
Chun-qiong SONG ; Hong-xing ZHUANG ; Shu-jie WANG ; Xiao-gen HU ; Le-ren HE
Chinese Journal of Plastic Surgery 2006;22(6):427-429
OBJECTIVETo investigate the possibility of external ear reconstruction using expanded postauricular scar flap and Medpor framework in burn cases.
METHODSExternal ear reconstruction using expanded postauricular scar flap in combination with Medpor framework was performed in 17 cases whose ear had burn injury.
RESULTSOf the 17 cases, 15 cases achieved success; 2 cases experienced partial exposure of the framework due to inadequate wrapping of the subcutaneous fascia flap and later injury. The longest follow-up was three years, and the final result was satisfactory.
CONCLUSIONSExternal ear reconstruction using expanded postauricular scar flap in combination with Medpor framework is a reliable method for adult (over 25 years) who has ear defect from burn injury.
Adult ; Burns ; complications ; surgery ; Cicatrix ; etiology ; surgery ; Ear, External ; injuries ; surgery ; Fascia ; transplantation ; Female ; Humans ; Male ; Middle Aged ; Polyethylenes ; Prosthesis Implantation ; Reconstructive Surgical Procedures ; methods ; Reoperation ; Stents ; Surgical Flaps
6.Identification of genes regulated by HBV preS1-transactivated protein 2 binding protein 1.
Huan-hu ZHANG ; Dan-qiong WANG ; Jun CHENG ; Jie ZHAO ; Xin-kai ZHAO ; Hong GAO
Chinese Journal of Hepatology 2010;18(4):267-270
OBJECTIVETo identify genes regulated by HBV preS1-transactivated protein 2 binding protein 1 (PS1TP2BP1).
METHODSPS1TP2BP1 gene was amplified by polymerase chain reaction (PCR) technique and cloned into the eukaryotic expression vector pcDNA 3.1/my-c-His A. The mRNAs isolated from HepG2 cells transfected recombinant eukaryotic expression vector pcDNA 3.1/myc-HisA-PS1TP2BP1 and pcDNA 3.1/myc-HisA empty vector were used to construct subtractive library. The differentially expressed genes were identified and analyzed.
RESULTS35 differentially expressed clones were obtained. Colony PCR identified 15 clones with 200-1000 bp inserts. Sequence analysis identified 15 differentially expressed genes.
CONCLUSIONThis study provides data for further characterize the function of PS1TP2BP1.
Amino Acid Sequence ; Base Sequence ; Carrier Proteins ; Cloning, Molecular ; Gene Library ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; Humans ; Molecular Sequence Data ; Protein Precursors ; genetics ; Trans-Activators ; genetics
7.Effect of nano-TiO(2) intratracheal instillation on lipid metabolism of AopE gene-knockout mice.
Jie-qiong HU ; Chun-ying CHEN ; Ru BAI ; Sen ZHEN ; Xian-mei DU ; Jia-jie ZANG ; Jiu-cun LI ; Yi-qun GU ; Guang JIA
Chinese Journal of Preventive Medicine 2010;44(9):780-784
OBJECTIVETo investigate the effect of nano-TiO(2) intratracheal instillation on the progression of dyslipidemia and atherosclerosis in apolipoprotein E-knockout mice.
METHODSThe nano-TiO(2) was ultrasound with phosphate-buffered saline solutions (PBS) into its suspension for exposure. A total of 46 specific pathogen free (SPF) level of 11-week-old male apolipoprotein E-knockout mice were randomly divided into groups by their body weights: non-treatment group (8 mice), PBS control group (9 mice), high dose group (1.0 mg/ml, 10 mice), medium dose group (0.5 mg/ml, 10 mice), and low dose group (0.1 mg/ml, 9 mice). Except the non-treatment group, mice from other groups were intratracheally instilled with 0.05 ml each time, twice a week. After exposure of 6 weeks, viscera index, blood TC, TG, HDL-C, LDL-C, and organic lipid ratio were assessed as biomarkers. Artery and aortic root issues were assessed by histopathology.
RESULTSAfter 5 weeks exposure, mice body weights in high dose group ((29.7 ± 1.9) g) started to drop, compared to PBS control ((31.3 ± 1.9) g, t = -1.58, P < 0.05) and low dose group ((31.4 ± 1.4) g, t = -1.17, P < 0.05); after 6 weeks, high dose group ((28.8 ± 1.5) g) was lower than PBS control ((30.4 ± 1.9) g, t = -1.60, P < 0.05), non-treatment group ((30.2 ± 1.3) g, t = -1.43, P < 0.05) and low dose group ((30.6 ± 1.0) g, t = -1.83, P < 0.05). TC levels of non-treatment, PBS control, high dose group, medium dose group and low dose group were (2.92 ± 1.18), (3.12 ± 0.73), (4.19 ± 1.86), (3.46 ± 0.72) and (2.57 ± 0.64) mmol/L, respectively; TG levels were (0.39 ± 0.13), (0.39 ± 0.08), (0.60 ± 0.21), (0.55 ± 0.19) and (0.41 ± 0.11) mmol/L, respectively; HDL-C levels were (1.67 ± 0.45), (1.54 ± 0.67), (0.93 ± 0.50), (1.02 ± 0.48) and (1.31 ± 0.64) mmol/L; TG levels of high dose group were higher than that of non-treatment group (t = 1.27, P = 0.03) and low dose group (t = 1.62, P = 0.01); TG levels of medium dose group was higher than PBS control (t = 0.16, P = 0.04), and TC levels of high dose group were higher than PBS control (t = 0.22, P = 0.01), non-treatment group (t = 0.22, P = 0.04) and low dose group (t = 0.20, P = 0.03), and HDL-C levels of high dose group were lower than PBS control (t = -0.61, P = 0.04) and non-treatment group (t = -0.74, P = 0.04); organic lipid ratio of each group were (2.27 ± 0.51)%, (2.06 ± 0.53)%, (2.90 ± 0.50)%, (2.60 ± 0.23)%, (2.24 ± 0.45)%; high dose group were higher than PBS control (t = 0.85, P = 0.00), non-treatment group (t = 0.64, P = 0.03) and low dose group (t = 0.67, P = 0.01); medium dose group was higher than PBS control (t = 0.54, P = 0.02). The plaque lipid content and calcium content which showed the progression of atherosclerosis and plaque rupture were elevated in medium and high dose groups.
CONCLUSIONIntratracheal instillation of nano-TiO(2) can induce dyslipidemia and accelerate the development of atherosclerosis and plaque rupture in ApoE-/-mice.
Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; blood ; chemically induced ; Dyslipidemias ; blood ; chemically induced ; Instillation, Drug ; Lipid Metabolism ; Lipids ; blood ; Male ; Mice ; Mice, Knockout ; Nanoparticles ; Specific Pathogen-Free Organisms ; Titanium ; administration & dosage ; pharmacology
8.IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis with its expression associated with DNA hypomethylation of exon 1.
Wen-jing RUAN ; Jie LIN ; En-ping XU ; Fang-ying XU ; Yu MA ; Hong DENG ; Qiong HUANG ; Bing-jian LV ; Hu HU ; Jing CUI ; Mei-juan DI ; Jian-kang DONG ; Mao-de LAI
Journal of Zhejiang University. Science. B 2006;7(11):929-932
Insulin-like growth factor binding-protein-7 (IGFBP7) was obtained from our previous colonic adenocarcinoma (CRC) and normal mucosa suppression subtraction hybridization (SSH) cDNA libraries. By RT-PCR and immunohistochemistry, we found that IGFBP7 was overexpressed in CRC tissue compared to normal tissue. However, our in vitro experiments performed in 10 CRC cell lines showed that IGFBP7 expressed only in SW480 and Caco2 cell lines, which implied an underlying reversible regulatory mechanism. Using methylation-specific PCR (MSP) and bisulfite sodium PCR (BSP), we found that its expression was associated with DNA hypomethylation of exon1. This was further supported by the in vitro study which showed restored IGFBP7 expression after demethylation agent 5-aza-2'-deoxycytidine treatment. Correlation analysis between IGFBP7 expression and prognosis indicated that overexpression of IGFBP7 in CRC tissue correlated with favourable survival. Investigation of the functional role of IGFBP7 through transfection studies showed that IGFBP7 protein could inhibit growth rate, decrease colony formation activity, and induce apoptosis in RKO and SW620 cells, suggesting it a potential tumor suppressor protein in colorectal carcinogenesis. In conclusion, our study clearly demonstrated that IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis and its expression is associated with DNA hypomethylation of exon 1.
Adenocarcinoma
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genetics
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metabolism
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Apoptosis
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genetics
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Cell Line, Tumor
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Colorectal Neoplasms
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genetics
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metabolism
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DNA Methylation
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Exons
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Gene Expression Regulation, Neoplastic
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genetics
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Humans
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Insulin-Like Growth Factor Binding Proteins
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genetics
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metabolism
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Transfection
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Tumor Suppressor Proteins
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genetics
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metabolism
9.Selection and quantitative detection of target genes in oral squamous cell carcinoma.
Li-qiong DUAN ; Wan-tao CHEN ; Ming-bin ZHANG ; Yong-jie HU ; Yan LÜ ; Ming YAN
Chinese Journal of Stomatology 2006;41(8):456-460
OBJECTIVETo select and identify the target genes related to oral squamous cell carcinoma (OSCC) and provide target genes for designing oligo-nucleotide functional microarray of OSCC.
METHODSGenes possibly related to oral squamous cell carcinoma were selected from the 5 years' published data of differently expressed profiles with microarray testing in OSCC. Then mRNA expression of selected genes were evaluated by real time quantitative polymerase chain reaction (RT-PCR) in 22 cases of OSCC, including tumor tissues and paried normal mucosas and quantified according to an internal control GAPDH.
RESULTSEight genes were tested. The overexpression of SPARC, PDGF-A, SERPINE1, TGF-beta(1) and VEGF-C genes were measured in 16, 18, 16, 20, 18 cases of tumor specimens, respectively. The expression of CK15 gene was lower than that of its normal tissue. There were overexpression of CCND1, BIRC3 in tumor tissues, but there was no significant difference of CCND1 and BIRC3 expression between tumor tissue and normal tissue (P > 0.05).
CONCLUSIONSSPARC, PDGF-A, SERPINE1, TGF-beta(1), VEGF-C and CK15 genes were closely related to tumor progress of OSCC. They can be used as the target genes for designing oligo-nucleotide functional microarray of OSCC.
Adult ; Aged ; Biomarkers, Tumor ; genetics ; Carcinoma, Squamous Cell ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mouth Neoplasms ; genetics ; Oligonucleotide Array Sequence Analysis ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods
10.Altered surfactant protein A gene expression and protein homeostasis in rats with emphysematous changes.
Qiong-jie HU ; Sheng-dao XIONG ; Hui-lan ZHANG ; Xue-mei SHI ; Yong-jian XU ; Zhen-xiang ZHANG ; Guo-hua ZHEN ; Jian-ping ZHAO
Chinese Medical Journal 2008;121(13):1177-1182
BACKGROUNDThe decrease of surfactant protein (SP) secreted by the alveolar type II cell is one of the important causes of limiting air of pulmonary emphysema. However, the SP-A gene and protein changes in this disease are rarely studied. This study was undertaken to investigate alterations in SP-A gene activity and protein, and to explore their roles in the pathogenesis of emphysematous changes.
METHODSTwenty Wistar rats were divided randomly into a normal control group (n = 10) and a cigarette smoking (CS) + lipopolysaccharide (LPS) group (n = 10). Ultra-structural changes were observed under an electron microscope. The number of cells positive for SP-A was measured by immunohistochemistry. The mRNA expression and protein level of SP-A in the lung tissues were determined by quantitative polymerase chain reaction (qPCR) and Western blot separately. The protein level of SP-A in lavage fluid was determined by Western blot.
RESULTSThe number of cells positive for SP-A of the CS + LPS group (0.35 +/- 0.03) was lower than that of the blank control group (0.72 +/- 0.06, P < 0.05). The level of SP-A in the lung tissues of rats in the CS + LPS group (0.2765 +/- 0.0890) was lower than that in the blank control group (0.6875 +/- 0.1578, P < 0.05). The level of SP-A in the lavage fluid of rats in the CS + LPS group (0.8567 +/- 0.1458) was lower than that in the blank control group (1.3541 +/- 0.2475, P < 0.05). The lung tissues of rats in the CS + LPS group showed an approximate increase (0.4-fold) in SP-A mRNA levels relative to beta-actin mRNA (P < 0.05).
CONCLUSIONSThe changes of SP-A may be related to emphysematous changes in the lung. And cigarette smoke and LPS alter lung SP-A gene activity and protein homeostasis.
Animals ; Blotting, Western ; Emphysema ; metabolism ; pathology ; Homeostasis ; Immunohistochemistry ; Male ; Microscopy, Electron ; Polymerase Chain Reaction ; Pulmonary Surfactant-Associated Protein A ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar