1.Molecular pathological changes of human fetal scleral fibroblasts following exposure to extremely low frequency electromagnetic field
Jie, WANG ; Jie-feng, CUI ; Huang, ZHU ; Li-quan, ZHAO ; Jie-yu, FENG ; Sheng-long, YE
Chinese Journal of Experimental Ophthalmology 2012;30(5):403-407
BackgroundThe effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood.ObjectiveThe present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.MethodsHFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining.Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased.ConclusionsELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.
2.High Selective Synthesis of 11?-hydroxycanrenone by Biotransformation
Rong-Sheng TAO ; Hai-Feng HU ; Xiao-Dun LI ; Bao-Quan ZHU ;
Microbiology 1992;0(02):-
Canrenone is an important intermediate for the synthesis of eplerenone,a cardiovascular drug.C_ 11 ?-hydroxylation of canrenone is the key reaction,which can be done by microbial transformation.Rhizopus sp.SIPI-0602,kept in our lab,could high selectively transform canrenone to a compound named SIPI-11.By determining and analyzing the MS,UV,NMR etc.spectra of compound SIPI-11,its chemical structure was elucidated to be 11?-hydroxycanrenone.The study on flask transformation technology showed that the transformation ratio exceeded 90% when the substrate concentration was not more than 6g/L.
3.SCREENING MICROBIAL HERBICIDES FROM WEED DRB
Han-Bin DAN ; Yong-Qiang CHEN ; Xue-Sheng WEI ; Zhu ZHU ; Liu TANG ; Yong-Quan LI ; Xiao-Feng ZHUANG ;
Microbiology 1992;0(04):-
By using enrichment media NPC and CVP, 792 strains of Pseudomonas and 515 strains of Erwinia were isolated from the rhizosphere of Digitaria adscendens (H. B. K.) Hem and Setaria vindis (L.) Beanv. Following which, experiments of antimetabolic test with E. coli, seed emergence controlling of S. viridis, herbicidal activity and security with green grass were carried out to select the desired bacteria. As the result, the selected strain, S7, could wholly control the seed emergence of S. viridis without any harm to the two tested green grass. And more, S7 promoted the seed emergence of Festuca arundinacea slightly. In spite of the comparatively low corrected mortality (56. 7%) of S7 after emergence of S. viridis, Selecting of microbial herbicides from weed DRB is thought to be more prospective.
4.Reverse second dorsal metatarsal artery island flap for repairing the soft tissue defect at toes.
Yu-feng JIAO ; Hai-quan WANG ; Yao-sheng LI
Chinese Journal of Plastic Surgery 2009;25(6):427-429
OBJECTIVETo report the application of reverse second dorsal metatarsal artery island flap for From May 2005 to September 2008, 5 cases with soft tissue repairing the soft tissue defect at toes.
METHODSdefects at toes were treated with reverse second dorsal metatarsal artery island flaps. The flaps size ranged from 2 cm x 3 cm to 5 cm x 6 cm.
RESULTSAll the 5 flaps survived completely. The patients could walk 1-2 months after operation. The patients were followed up for 5-7 months with good appearance, texture and sensation of toes.
CONCLUSIONThe reverse second dorsal metatarsal artery island flap has a reliable blood supply and good tissue texture. It is a practical method for repairing the soft tissue defect at toes.
Adult ; Fascia ; transplantation ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Toes ; injuries ; Young Adult
5.A quantitative study on ultrasonographic texture analysis of injured porcine striped muscle induced by thermal and chemical factors
Jia-qi, ZHAO ; Jian-quan, ZHANG ; Qi, XU ; Yan-qiu, CHEN ; Jian-guo, SHENG ; Feng, LU ; Tian, YANG
Chinese Journal of Medical Ultrasound (Electronic Edition) 2013;(8):674-680
Objective To quantitatively analyze and compare the texture features of thermal and chemical lesions on the porcine striated muscle, in vitro extracted from high-frequency ultrasonograms using computer-assisted image analysis technique, and to investigate the application values. Methods The thermal lesion and chemical lesion were induced in vitro in porcine striated muscle by microwave ablation and anhydrous acetic acid injection, respectively. The two dimension (2D) ultrasonographic ifndings were qualitatively compared between the groups of thermal and chemical lesion models, in which eight textural features in geometric mathematics extracted from 2D ultrasonograms were quantitatively analyzed by a technique of computer-assisted image analysis named multiscale decomposition method of echo intensity of interface relfections. Results As expected, microwave ablation and anhydrous acetic acid caused signiifcant changes of several texture features extracted from ultrasonograms. There were significant differences between the normal group and microwave ablation group in grayscale mean (Mean), irregularity (IRGL) and periodicity of distribution (POD) as follows (Mean: 1.9143±0.2914 vs 1.2334±0.3357, t=-5.306, P=0.000; IRGL: 0.5577±0.0334 vs 0.5092±0.0459, t=-2.957, P=0.007; POD: 0.000 27±0.000 005 vs 0.000 29±0.000 008, t=4.782, P=0.000). There were signiifcant differences between the normal group and anhydrous acetic acid injection group in number of blobs (NOB), size of blobs (SOB) and periodicity of distribution (POD) as follows (NOB: 51.0324±13.6998 vs 31.6042±4.8315, t=4.633, P=0.000; SOB:16.4843±3.9349 vs 25.6230±2.3555, t=6.903, P=0.000;POD:0.000 26±0.000 015 vs 0.000 29±0.000 008, t=-4.459, P=0.000). For each group of injured regions, there were significant differences between the microwave ablation group and anhydrous acetic acid injection group in Mean, IRGL, NOB and SOB as follows (Mean: 1.2664±0.2688 vs 1.9143±0.2914, t=-5.661, P=0.000; IRGL: 0.5220±0.0422 vs 0.5577±0.0334, t=-2.295, P=0.032;NOB:51.0324±13.6998 vs 34.5856±2.6362, t=4.048, P=0.000;SOB:16.4843±3.9349 vs 25.3176±2.3501, t=-6.676, P=0.000). Conclusion Technique of computer-assisted image analysis named multiscale decomposition method of echo intensity of interface relfections, based on multiscale blob features extraction, was useful to differentiate ultrasonic texture features between the groups injured in our study, which established quantitative muscle ultrasound as a practical and reliable tool for the muscle injury diagnosis to distinguish the structural changes induced by different physiochemical factors.
6.Influence of fluoride on osteoprotegerin mRNA and protein expression in rat osteoblasts: an in vitro study
Dan, LI ; Xing-quan, JIANG ; Xiao-feng, JI ; Zhi-tao, ZHAO ; Zhe, FAN ; Ling, JING ; Guang-sheng, LI
Chinese Journal of Endemiology 2009;28(2):134-137
Objective To study the influence of fluoride on the expression of osteoprotegerin(OPG) mRNA and protein in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarial of suckling Wistar rats were cultured in vitro in the media supplemented with NaF at a series of doses[O(control), 1,2 and 4 mg/L groups], and OPG mRNA expression and protein were evaluated by RT-PCR and ELISA methods, respectively. Results OPG mRNA expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h and 72 h(F=333.48,808.34,P<0.05). OPG mRNA expression in suckling rat osteoblasts cultured in vitro after exposure to NaF for 48 h at different doses(0.810±0.003, 0.819±0.031 and 0.870±0.044 for 1,2 and 4 mg/L groups, respectively) compared with that of control (0.800±0.040, all P<0.05). OPG mRNA expression further increased for 72 h exposure to NaF(0.933±0.047,1.031±0.051,1.240±0.062 for 1,2 and 4 mg/L, respectively), significantly higher than that of the control (0.805±0.020,all P<0.05) and corresponding groups at 48 h. NaF doses and time exposure exhibited a significant synergistic effect on OPG mRNA expression(F=2004.16, P<0.05). NaF also enhanced OPG protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed only in 4 mg/L group(0.228±0.014,0.277±0.048) and control(0.205±0.012,0.229±0.010) at 48 h and 72 h (P<0.05). In addition, OPG protein expression at 72 h post-exposure was higher than that at 48 h,but there was no synergistic effect between concentration and time(F=1.21,P>0.05). Conclusions The results suggested that NaF could increase OPG mRNA and protein expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.
7.Effects of trimetazidine on myocardial ischemia and heart rate variability in patients with non ST-elevation acute coronary syndrome
Jing ZHANG ; Sheng-Hu HE ; Shu CHEN ; Yi ZHANG ; Feng-Di YAN ; Jian-Feng YAN ; Fu-Quan ZHAO ; Xiao-Hua XU ;
Chinese Journal of Clinical Pharmacology and Therapeutics 2004;0(12):-
AIM:To observe the effects of trimetazi- dine on myocardial ischemia and heart rate variability (HRV)in patients with non ST-elevation acute coronary syndrome(NSTEACS).METHODS:74 patients with NSTEACS were chosen and randomly assigned into two groups:one group was given conventional therapy with as- pirin,isosorbide mononitfate,fluvastatin and metoprolol, trimetazidine was added on the base of conventional thera- py in the other group.The total observation time of thera- py was 8 weeks.The heart rote,blood pressure,frequen- cy and persistence time of chest pain,numbers of times of using Nitroglycerin every day,the extent anti degree of myocardial isehemia,HRV of patients were observed be- fore and after treatment.RESULTS:The therapeutic ef- fect of trimetazidine treatment group(with the effective rate 94.4%,excellence rate 72.2%)was obviously bet- ter than that of the conventional treatment group(with the effective rate 65.8%,excellence rate 36.8%)(P
8.The experimental study on porous calcium phosphate cement with bone marrow stromal cells for bone tissue engineering.
Lei WANG ; Yao-jun LI ; Yan ZHANG ; Ke-feng PAN ; Yuan-liang HUANG ; Chang-sheng LIU ; Xin-quan JIANG
West China Journal of Stomatology 2010;28(3):315-318
OBJECTIVETo observe the biocompatibility of new biomaterials porous calcium phosphate (CPC) and ectopic bone formation of CPC with bone marrow stromal cells (BMSCs).
METHODSThe BMSCs were cultured from Beagle dog and combined with the porous CPC with the best concentration after transfect green fluorescent protein (GFP). The adhesion and growth of BMSCs on CPC were observed under inversion, fluorescence and scanning electron microscopy. The ectopic bone formation were observed at the 8th week after CPC and BMSCs were implanted subcutaneously into nude mice.
RESULTSWhen BMSCs with CPC were cultured at the 1st day, cells were climbing out from CPC with normal morphology. At the 7th day cells can be seen protruding pseudopods, secretion of matrix. Bone formation could be seen histomorphologically at the 8th week.
CONCLUSIONPorous CPC has good biocompatibility and is an ideal scaffold material for bone tissue engineering.
Animals ; Biocompatible Materials ; Bone Cements ; Bone and Bones ; Calcium Phosphates ; Dental Cementum ; Dogs ; Mesenchymal Stromal Cells ; Mice ; Mice, Nude ; Tissue Engineering
9.A preliminary study about the apoptostic mechanism of RNA targeting basic fibroblast growth factor in glioma U251 cells.
Jing-chao WU ; Xue-quan FENG ; Jin-huan WANG ; Jun LIU ; Biao ZHANG ; Xin-nü XU ; Hong-sheng LIU
Chinese Journal of Surgery 2012;50(10):928-932
OBJECTIVETo preliminarily investigate the mechanism of small interfering RNA (siRNA) induced apoptosis in glioma U251 cells by silencing basic fibroblast growth factor (bFGF).
METHODSU251 cells were divided into the normal control group, the mock group and experiment group, the mock and experiment group were transfected with mock vector (Ad-null) and the recombinant adenovirus carrying bFGF-siRNA (Ad-bFGF-siRNA) respectively at a multiplicity of infection (MOI) of 100. After 72 hours, the expression of related proteins was revealed by the method of Western blot. Mitochondrial transmembrane potential (ΔΨm) was measured with flow cytometry and confocal microscopy, Groups were compared using single factor analysis of variance (One-way ANOVA).
RESULTSAfter U251 cells were transfected with bFGF-siRNA, the results of Western blot showed that after 72 hours of transfection the bFGF protein in the experiment group decreased obviously, meanwhile Cytochrome C, Caspase-3 and Bax showed increased expression while in the Bcl-xl and Bcl-2 proteins decreased expression. The proportion of high mitochondrial membrane potential of cells by flow cytometry, the experimental group was 74.4% ± 4.7% decreased significantly compared with the control group 92.1% ± 2.5%, the mock group 90.9% ± 1.8% (F = 28.805, P < 0.05); laser scanning confocal microscopy results showed that the red fluorescence and green fluorescence ratio of the experimental group was 0.83 ± 0.12 decreased significantly compared with 1.36 ± 0.40 of the control group and 1.32 ± 0.35 of the mock group(F = 7.920, P < 0.05).
CONCLUSIONsiRNA targeting bFGF induced U251 cell apoptosis may be achieved through the mitochondrial pathway.
Adenoviridae ; genetics ; Apoptosis ; Cell Line, Tumor ; Fibroblast Growth Factor 2 ; genetics ; Glioma ; pathology ; Humans ; RNA Interference ; RNA, Small Interfering ; Transfection
10.Optimization of culture measure for bovine-bovine and goat-bovine cloned embryos in vitro.
Lin ZHANG ; Song HUA ; Yong ZHANG ; Fu-Sheng QUAN ; Feng-Jun LIU ; Lie-Ru LIAO ; Yong-Hai JIANG
Chinese Journal of Biotechnology 2007;23(4):662-666
UNLABELLEDThis study is conducted to explore an effective culture method for supporting the embryo development. The cattle fetal ear fibroblasts and the goat fetal ear fibroblasts are transplanted into the enucleated cattle oocytes separately by oocyte intraplasmic nuclear injection method to construct bovine cloned embryos and goat-bovine cloned embryos. The embryos are first cultivated in modified charles rosenkrans 2 amino acid medium (mCR2aa) and modified synthetic oviduct fluid medium (mSOF) separately. Then BSA (8 mg/mL) or FBS (10%) can be added to mSOF according to the different culture period. The supplements and orders, added during the first three days and after three days are as follow: BSA and BSA, BSA and FBS, FBS and BSA, FBS and FBS. On the basis of the cleavage rate, 8/16-cell rate, blastocysts rate and total cell number of blastocysts, the best culture way can be screened out.
RESULTFirst, cleavage rate, 8/16-cell rate, blastocysts rate and total cell number of blastocysts, cultivated in mSOF solution are all higher than those cultivated in mCR2aa( P < 0.05). Second, the cleavage rate and 8/16-cell rate, adding BSA and FBS into mSOF, are in turn 79.8% +/- 7.1%, 49.7% +/- 3.5%, 21.5% +/- 1.8%, and 115.2 +/- 4.3 in bovine cloned embryo, and 40.1% +/- 6.3%, 29.2% +/- 2.0%, 13.4% +/- 2.1% and 100.1 +/- 3.0 in goat-bovine cloned embryo, which are significant higher than other culture groups (P < 0.05).
CONCLUSIONThe goat-bovine cloned embryo can be cultivated by the optimized culture measure of bovine cloned embryo. The best culture ways of bovine cloned embryo and goat-bovine cloned embryo are all to use mSOF supplemented BSA in the first three days and then use mSOF supplemented FBS in the next five days.
Animals ; Cattle ; embryology ; physiology ; Cells, Cultured ; Cloning, Organism ; veterinary ; Ear, External ; cytology ; Embryo Culture Techniques ; methods ; veterinary ; Embryonic Development ; Fibroblasts ; cytology ; transplantation ; Goats ; embryology ; physiology ; Nuclear Transfer Techniques ; Oocytes ; cytology