1.Action mechanism of narrow-band ultraviolet B on vascular regulatory factors in psoriasis vulgaris
Fumin LI ; Xue WANG ; Xiling DUAN ; Linhong SONG ; Dandan DONG ; Wei DONG ; Xun HE ; Chuanpeng YING
Chinese Journal of Dermatology 2009;42(3):163-166
Objective To evaluate the influence of narrow-band ultraviolet B on lesional microvessel density (MVD),vascular endothelial growth factor (VEGF),matrix metalloproteinases 2 (MMP-2)as well as on serum VEGF in patients with psoriasis vulgaris(PV).Methods Fifteen patients with PV were recruited into this study with 10 normal human controls.All patients received NB-UVB phototherapy thrice a week for 4-5 weeks.Prior and after the treatment,psoriasis area and severity index (PASI)was calculated,tissue specimens were taken from non-photoexposed lesions,and sera samples were obtained from these patients.Then,MvD and the expression level of VEGF and MMP-2 were measured by immunohistochemical labeled dextran polymer(LDP)method in the tissue specimens.Also,the serum level of VEGF was tested by enzyme-linked immunosorbent assay(ELISA).Results PASI score remarkably decreased in patients after the photothempy(t=13.35,P<0.01).The MVDs were 20.52±5.02,7.33±1.24 and 4.26±0.79 capillaries per high power field(400 × amplification),in psoriatic lesions before treatment,after treatment,and normal control tissues,respectively,with a significant difference among the three groups (F=97.57,P<0.05),and a significant increase was observed in the lesions before treatment compared with those after treatment and normal controls.The serum level of VEGF was 307.55±121.65 ng/L in psoriatic lesions before treatment,significantly higher than that after treatment(163.92±95.57 ng/L),and in normal control skin (139.78±79.06 ng/L),whereas there was no significant difference between the latter two groups(P>0.05).The positivity rate of MMP-2 was similar among the three groups without statistical difference(P>0.05).In psoriatic patients,a positive correlation was observed among PASI score,MVD,lesional and serum VEGF levels(P<0.05),also among the MVD,VEGF and MMP-2 levels in lesions(P<0.05).but lesional MMP-2 was unrelated to PASI score or sgrum VEGF(both P>0.05).Conclusions NB-UVB may regulate superficial dermal microvascular proliferation by acting on the expression of VEGF in sera and lesions of psoriatic patients.VEGF and MMP-2 may bOth participate in the proliferation process of microvessels,while MMP-2 is unlikely to be involved in the therapeutic mechanism of NB-UVB.
2.Cloning and Characterization of 5′Flanking Region of Nitrate Reductase Gene Derived from Dunaliella salina
Jie LI ; Yan-Long JIA ; Hong-Xia YAN ; Wei-Dong PAN ; Le-Xun XUE ;
China Biotechnology 2006;0(11):-
Aim: Clone and characterize of the 5′- flanking region of the nitrate reductase (NR) gene derived from Dunaliella salina(D. salina). Methods : The genomic DNA from D. salina was respectively digested with BamHI, EcoRI, HindIII, Pst I, Sal I and Xba I. A genomic walking cassette was ligated to the ends of the digested DNA fragments, and then genomic walking libraries comprising BL, EL, HL, PL, SL and XL were constsucted. The 5′- flanking region of the NR gene from genomic walking libraries of D. salina was amplified by LA-PCR. The DNA sequences were analyzed with the software - Promoter Predictions. Isolated 5′-flanking regions fused to the GUS gene were tested for transient expression in the alga. Results: A single specific PCR product of about 1200bp in length from the HL library was generated. Also, several conserved motifs, such as CAAT-box, GAGA-box were found, which are related to regulation of transcription, and the putative binding sites of transcriptional factors such as EBP, EFII, NF-E1 and LV. BLAST showed that the DNA sequences shared high homology with 5′-upstream region of the NR gene from Dunaliella viridis. The isolated 5′-flanking regions were able to strongly drive GUS reporter gene expression, suggesting that it contains the promoter elements necessary for the transcription of the NR gene. The expression pattern of the GUS gene and the NR gene were similar, both ware induced by nitrate and repressed by ammonium. Conclusion: The cloned 5′- flanking sequences of NR gene derived from D. salina might be a specific promoter with the ability to“switch on or off” an expression of the heterologous gene in transgenic D. salina.
3.Long-term efficacy comparison between goniosynechialysis and combination of phacoemulsification with goniosynechialysis for the management of chronic angle closure glaucoma
Jing, ZHU ; Wei, ZHAO ; Jun, SHAO ; Xun, BAO ; Jing, LIN ; Dong-hong, FU ; Yong, YAO
Chinese Journal of Experimental Ophthalmology 2013;31(10):964-967
Background One of the features of the pathogenesis of primary angle-closure glaucoma(PACG) is anterior synechia of peripheral iris.Goniosynechialysis and combination of phacoemulsification and goniosynechialysis have been applied for the treatment of the disease recently,but the selection of operative types has great impact on clinical efficacy.Objective This study was to investigate the long-term efficacy of goniosynechialysis and combination of phacoemulsification and goniosynechialysis for the management of chronic PACG.Methods A non-randomized clinical controlled trial was designed.This clinical trial complied with Declaration of Helsinki and was approved by Medical Ethic Committee of Nanjing Medical University.Written informed consent was obtained from each patient.One hundred and ten eyes of 110 patients with chronic PACG were assigned to the goniosynechialysis group and combined operative group from March,2008 to February,2011 in Wuxi People's Hospital.180° goniosynechialysis was performed on 34 patients in the goniosynechialysis group,and phacoemulsifieation +intraocular lens (IOL) implantation + goniosynechialysis were carried out in 78 patients of the combination operative group.All the patients were followed-up for 2 years.Vision acuity,intraocular pressure (IOP),anterior chamber depth(ACD),unltrasound biomicroscopy and perimetry were recorded and compared between before and after operation.Results No significant difference was found in vision acuity between preoperation and postoperative 2 years in the goniosynechialysis group ([0.65 ± 0.15] vs.[0.45 ± 0.15]) (t =1.57,P>0.05),but in the combination operative group,the vision acuity was significantly different between the before and after operation ([0.25±0.15] vs.[0.85 ±0.05]) (t =9.12,P<0.001).The lOPs at 2 years after operation were (14.2±4.1) mmHgand(13.7±4.8) mmHg,respectively in the goniosynechialysis group and combination operative group and were significantly lower than(47.2 ±6.3) mmHg and(46.9±7.0) mmHg before operation(t =4.95,P<0.001 ;t=5.03,P<0.001).The ACD values in the goniosynechialysis group and combination operative group were(3.38±0.02)mm and (3.54±0.03) mm 2 years after operation,which were significantly increased in comparison with (1.33 ±0.24)mm and (1.56±0.37) mm before operation(t=7.65,P<0.001;t=6.76,P<0.001).Conclusions Both combination of phacomulsification with goniosynechialysis or 180° goniosynechialysis are effective for the treatment of PACG.Suitable operation should be alternated depending on the indicators of PACG patients.
4.Profiling Membrane Proteome of Macrophages by One-dimensional PAGE and Liquid Chromatography-Tandem Mass Spectrometry
Ling-Bing ZHANG ; Yan-Ni LUN ; Le-Yang YU ; Dong-Mei YAN ; Wei MA ; Bai-Rong DU ; Xun ZHU ;
China Biotechnology 2006;0(04):-
Macrophages are involved in many important biological processes and membrane proteins are the key effector molecules for their functions. However, membrane proteins are difficult to analyze by 2-DE based method because of their intrinsic tendency to self-aggregate during the first dimension separation (IEF). To circumvent the obstacle hampering membrane protein analysis, we combined one-dimensional SDS-PAGE with capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using this technique, we identified 458 GO annotated membrane proteins with extremely high confidence, including most known markers of peritoneal macrophages (e.g., CD11b, F4/80, CD14, CD18, CD86, CD44, CD16 and Toll-like receptor). Thirteen other CD antigens and 18 Ras-related small GTPase were also identified. In addition to those known macrophage membrane proteins, a significant number of novel proteins have also been identified. This research provides a valuable data set of macrophage membrane proteins, thus allowing for more comprehensive study of membrane proteins and a better understanding of the function mechanisms of macrophages in many biological processes.
5.Intervention effect of quercetin on inflammatory secretion of cardiac fibroblasts.
Xi-Lan TANG ; Jian-Xun LIU ; Wei DONG ; Peng LI ; Lei LI ; Yong-Qiu ZHENG ; Jin-Cai HOU
China Journal of Chinese Materia Medica 2014;39(12):2314-2317
To establish neonatal rat cardiac fibroblast inflammatory secretion model by using LPS 100 microg x L(-1) combined with ATP 5 mmol x L(-1), in order to study the inhibitory effect of quercetin on the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts, further investigate the effect of quercetin on the protein expression of p-NF-kappaB p65 (S276) and p-Akt (S473) by western blot, and discuss the inhibitory effect of quercetin on the inflammatory secretion of cardiac fibroblasts. According to the findings, quercetin with the concentrations between 51.74 micromol x L(-1) and 827.81 micromol x L(-1) had no significant effect on the activity of cardiac fibroblasts. Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of TNF-alpha and IL-1beta induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.01 or P < 0.05). Quercetin with the concentrations of 82.78, 41.39 micromol x L(-1) could notably inhibit the increase of IL-6 induced LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.05), without any notable effect of quercetin with the concentration of 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20. 70 micromol x L(-1) could notably inhibit the NF-kappaB p65 (S276) activation induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 15 min, with the most significant effect in 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of p-Akt(473) expression induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 240 min (P < 0.05). Therefore, this study believes that quercetin could attenuate the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts by inhibiting the activation of NF-kappaB p65 (S276) and Akt (473).
Animals
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Cells, Cultured
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Drugs, Chinese Herbal
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administration & dosage
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Endomyocardial Fibrosis
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drug therapy
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genetics
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immunology
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Female
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Fibroblasts
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drug effects
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immunology
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Heart
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drug effects
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Humans
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Interleukin-6
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genetics
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immunology
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Male
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Proto-Oncogene Proteins c-akt
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genetics
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immunology
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Quercetin
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administration & dosage
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Rats
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Rats, Sprague-Dawley
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Tumor Necrosis Factor-alpha
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genetics
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immunology
6.Studies on properties of depotentiation of long-term potentiation induced by low frequency stimulation in CA1 neurons of rat hippocampal slices.
Hua-Wei LIANG ; Yue-Liang SHEN ; Xun-Dong WU
Chinese Journal of Applied Physiology 2002;18(3):218-221
AIM AND METHODSThe parameters of low frequency stimulation (LFS) were altered systematically (frequencies of 1, 3 or 5 Hz; number of pulses of pulses of 300 or 900; and time lag after high frequency stimulation (HFS) of 20 or 100 min) and examined their effects on depotentiation (DP) of long-term potentiation (LTP) of synaptic transmission in CA1 neurons in hippocampal slices of rat.
RESULTSLTP could be induced by HFS (two trains of 100 Hz, 100 pulses, separated by 30 s) and be reversed to produce DP by a train of LFS of 900 pulses at 3 Hz given 20 min after HFS. DP induced by LFS could be blocked by NMDA receptor antagonist AP5 (50 micromol/L). And significantly reduced effect was observed for LFS at 1 Hz or 5 Hz, with smaller numbers of pulses or a longer time lag from LFS to HFS.
CONCLUSIONThe above results indicate that DP induced in CA1 neurons of rat hippocampal slices is strongly dependent on the parameters of LFS, and the process may be mediated through the NMDA receptor.
Animals ; Electric Stimulation ; methods ; Hippocampus ; cytology ; physiology ; In Vitro Techniques ; Long-Term Potentiation ; Male ; Neurons ; physiology ; Rats ; Rats, Sprague-Dawley ; Synapses ; physiology ; Synaptic Transmission ; physiology
7.Observations on the Therapeutic Effect of"Neck Seven Needles"on Cervical Spondylotic Radiculopathy
Jie LI ; Wei XIE ; Wei-Qiang FAN ; Dong-Xun ZHU ; Hai-Bin ZHANG ; Hai-Ping HONG
Shanghai Journal of Acupuncture and Moxibustion 2018;37(2):222-225
Objective To investigate the clinical efficacy of Zhu's "neck seven needles" in treating cervical spondylotic radiculopathy. Method Ninety-six patients with cervical spondylotic radiculopathy were randomized to two groups. The treatment group received Zhu's "neck seven needles" therapy and the control group, cervical vertebra traction therapy. Both groups were treated once every other day, for a total of 10 days. The symptoms were scored using the Short-form McGill Pain Questionnaire (SF-MPQ-2) before and after treatment. Result The SF-MPQ-2 score decreased significantly in both groups of patients after treatment compared with before. It decreased more in the treatment group than in the control group and the difference was statistically significant (P<0.01). The total efficacy rate was significantly higher in the treatment group than in the control group (P<0.01). Conclusion Zhu's "neck seven needles" is clinically more effective than conventional cervical vertebra traction in treating cervical spondylotic radiculopathy.
8.Hypoxia Downregulates the Angiogenesis in Human Placenta via Notch1Signaling Pathway
LI YU-QI ; LIU HAI-YI ; CAO LAN-LAN ; WU YUAN-YUAN ; SHI XIN-WEI ; QIAO FU-YUAN ; FENG LING ; DENG DONG-RUI ; GONG XUN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):541-546
Placentation,which is critical for maternal-fetal exchange of nutrients and gases,is a complicated process comprising stepwise vasculogenesis and angiogenesis.Hypoxia caused by impairedtrophoblast invasion may cause various angiogenic abnormalities in human placenta.The Notchl signaling pathway plays an important role in the regulation of angiogenesis.The angiogenesis of human umbilical vein endothelial cells (HUVECs) under normal/hypoxic conditions and the mRNA/protein level of Notchl/Dell4/Jaggedl were investigated in this study.The effects of DAPT/JAG-1 on the migration of HUVECs were also assessed by cell wound healing assay,so as to discover the possible role of notchl signaling pathway in the angiogenesis of human placenta.The results showed that angiogenic ability of HUVECs was seriously reduced under hypoxic conditions.The mRNA and protein levels of Notchl/Dell4/Jaggedl were decreased in the hypoxic group compared to the control one.In addition,the migration capability of HUVECs was significantly obstructed when treated with DAPT and under hopoxic condition,but promoted when treated with JAG-1.The above results demonstrate that hypoxia downregulates the angiogenesis in human placenta via Notch 1 signaling pathway.
9.Balloon kyphoplasty:a view from points of technique and technique-related issues
Gang SUN ; Peng JIN ; Xun-Wei LIU ; Run-Song HAO ; Zhi-Yong XIE ; Fan-Dong LI ; Yu-Hai YI ; Xu-Ping ZHANG ;
Chinese Journal of Radiology 2000;0(11):-
Objective To discuss the technique and technique-related issues of percutaneous kyphoplasty(PKP).Methods The study involved 69 vertebrae in 51 cases of painful osteoporotic vertebral compressive fractures.Under X-ray fluoroscopy monitoring,the fractured vertebral bodies were treated by kyphonplasty with inflatable balloon.The preoperative and postoperative vertebral height and Cobb angle in radiography were measured and analyzed.Results All patients tolerated the procedure well with dramatic pain relief within 72 hours after the procedure.No clinical complication was found.The loss heights of the anterior and mid portion of the vertebral body reduced from 15?4mm and 11?4mm preoperatively to 10? 4mm and 6?3mm postoperatively,respectively.Cobb angle corrected averagely from 22??6? preoperatively to 12??4?.There was significant difference between preoperative and postoperative measures (P
10.Cloning and analysis of psaB cDNA of Dunaliella salina.
Hong-Tao LIU ; Wei-Dong ZANG ; Zhao-Ming LU ; Ning WANG ; Gui-Qin HOU ; Shen-Ke LI ; Le-Xun XUE
Chinese Journal of Biotechnology 2005;21(4):642-645
One pair of degenerate primer was designed according to conserved motifs of the psaB (A2 subunit of photosystem I) of Chlamydomonas reinhardtii, Chlamydomonas moewusii, Chlorella vulgaris and Mesostigma viride, and a total RNA of Dunaliella salina (D. salina) was extracted with TRIzol reagent. A cDNA fragment, about 1.8kb in length, from green algal D. salina was obtained through RT-PCR method. The resulting PCR product was cloned into T-vector and screened to determine its sequence. Homologous analysis of the deduced amino acid sequence was performed by BLAST and subsequeqtly compared with GenBank data. The obtained cDNA sequence was 1815 bp long, which encodes 605 amino acids (GenBank accession number: AY820754). The sequence shared high homologue with the following psaB: Chlamydomonas reinhardtii 92%, Chlamydomonas moewusii 91%, Chlorella vulgaris 86%, Mesostigma viride 85%, Physcomitrella patens subsp. Patens 85% and Nephroselmis olivacea 84%. It can be concluded that the cloned sequence is psaB cDNA fragment from D. salina.
Algal Proteins
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genetics
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Amino Acid Sequence
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Animals
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Chlamydomonas reinhardtii
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genetics
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Chlorophyta
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genetics
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metabolism
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Cloning, Molecular
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DNA, Complementary
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genetics
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Molecular Sequence Data
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Photosystem I Protein Complex
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genetics
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Sequence Analysis, Protein
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Sequence Homology, Amino Acid