1.Effects of interleukin-1 ,interleukin-6 and tumor necrosis factor-α on migration of human lens epithelial cells
Chinese Journal of Experimental Ophthalmology 2012;(10):873-876
Background Lens epithelial cells (LECs)play an important role in maintaining the lens transparency.Inflammatory cytokines have been clarified to participate in the formation of traumatic and after cataracts.However,the influence of inflammatory cytokines on the migration of LECs is still studying.Objective This study was to investigate the effects of interleukin-1 alpha (IL-1α),interleukin-1 beta (IL-lβ),IL-6,tumor necrosis factor alpha(TNF-α) and their mixture on the migration of human LECs in vitro.Methods HLE-B3 cells were cultured in DMEM containing 0.5% fetal bovine serum.10 μg/L of IL-1α,IL-1β,IL-6,TNF-α or the mixture was added in the medium for 24 hours,respectively.Wound-scratch assay and transwell monolayer permeability assay were used to evaluate the effects of different cytokines on LECs migration.The migrating cell numbers at the scratch zone and the transmembrane cell numbers at 12:00,3:00,6:00,9:00 and the center areas on the transwell chamber were calculated under the inverted microscope.In the control group,LECs were cultured in DMEM with 0.5% fetal bovine serum only.Results Wound-scratch assay revealed that 24 hours after cytokine incubation,the LECs numbers in the scratch zone were significantly less in the control group compared with the IL-1β group and TNF-αgroup (P =0.000,0.000),and those in the mixture group were higher than the IL-1β group and TNF-α group (P=O.000,0.000).However,no significant change was found in migrating LECs number between IL-1o group or IL-6 group and the control group (P =0.600,0.098).Transwell assay displayed that after 24-hour culture with cytokines,the largest density of transmembrane LECs was obtained in the mixture group,with a significant difference in comparison with the IL-1β group or TNF-α group (P=0.000,0.000).However,the transmembrane LECs were much more in the IL-1 β group or TNF-α group compared with the control group (P =0.000,0.000).Only a few transmembrane LECs were seen in the IL-1α group and IL-6 group in comparison with the control group(P =0.056,0.327).Conclusions IL-lα and IL-6 can not stimulate the migration of human LECs,but TNF-α and IL-1β can enhance their migration markedly.Mixture of these cytokines may play a much powerful influence on LECs migration.
2.Effect of the pathological changes of the pelvic cavity and fallopian tube on the outcome of in vitro fertilization and embryo transfer
Min ZHANG ; Hongyan ZOU ; Min DU ; Jian WANG ; Rongping YU
Chinese Journal of Postgraduates of Medicine 2011;34(21):17-19
Objective To evaluate the effect of the pathological changes of the pelvic cavity and fallopian tube on the outcome of in vitro fertilization and embryo transfer(IVF-ET).Method One thousand and thirty-two patients who underwent IVF-ET were divided into tubal and pelvic infertile group(605 cases)and non-tubal and pelvic infertile group(427 cases).The tubal and pelvic infertile group was also divided into salpingemphraxis group(243 cases),tubal resection group(104 cases),fallostomy group(149 cases),tubal dropsy group(109 cages)according to the tubal lesion regions,and combined with pelvic group(194 cases),combined without pelvic group(411 cases).The data of clinical pregnancy,ectopic pregnancy,and abortion was analyzed respectively.Results The ectopic pregnancy and abortion rates in tubal and pelvic infertile group[10.63%(27/254)and 9.06%(23/254)]were higher than those in non-tubal and pelvic infertile group [3.27%(5/153)and 4.58%(7/153)](P<0.01 or<0.05).The ectopic pregnancy rate was the lowest in tubal resection group[2.17%(1/46)],the highest in fallostomy group[22.41%(13/58)],there was significant difference among the groups(P<0.01).The abortion rate in fallostomy group and tubal dropsy group[10.34%(6/58)and 15.00%(6/40)]was higher than that in salpingemphraxis group and tubal resection group [7.27%(8/110)and 6.52%(3/46)],there was significant difference among the groups(P<0.05).The abortion rate in combined with pelvic group[11.54%(9/78)]was higher than that in combined without pelvic group[7.95%(14/176)](P<0.05).Conclusions The pathological changes of the pelvic cavity and fallopian tube are higher risk factors of ectopic pregnancy and abortion occurrence.The assessment and treatment of pelvic cavity and fallopian tube before assisted reproductive treatment cycles should be enhanced.
5.Research progress of pretreatment of biological samples.
Jian-nan FENG ; Shou-ying DU ; Jie BAI ; Yang LU ; Hui-min LIU
China Journal of Chinese Materia Medica 2014;39(21):4143-4148
Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals
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Body Fluids
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chemistry
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Chemical Precipitation
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Chemistry Techniques, Analytical
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methods
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Humans
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Liquid-Liquid Extraction
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Proteins
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isolation & purification
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Solid Phase Extraction
6.MECP 2 duplication syndrome:a pedigree report and literature review
Min ZHU ; Yue ZHANG ; Jian TANG ; Senjie DU ; Dalin FU ; Xiaoke ZHAO ; Hongying LI ; Li ZHANG
Journal of Clinical Pediatrics 2016;34(12):898-902
Objective To explore the clinical features and the gene mutations in MECP 2 duplication syndrome. Methods The clinical data of a child with developmental retardation and hypophrenia accompanied with respiratory tract infection was analyzed retrospectively. Microarray analysis technique was used to detect the genes in the patient and his family. The pertinent literature was reviewed. Results A 1-year and 7-month old boy was found to have hypotonia, developmental delay, and recurrent respiratory tract infections after birth. Microarray analysis showed a duplication of 441.88kb in Xq28 area and diagnosis of MECP2 duplication syndrome was confirmed. His grandmother, mother, and two aunts were found duplication of 441.73-441.88kb in Xq28 area, all of whom were MECP2’s female carrier. Conclusions The improvement of chromosome chip technology inspection is helpful to the early diagnosis of MECP2 duplication syndrome.
7.The novel surface-anchored protein SasX promotes aggregation and colonization of Staphylococcus aureus
Jian CHEN ; Xin DU ; Yan SONG ; Feiyi RUAN ; Yuan Lü ; Min LI
Chinese Journal of Microbiology and Immunology 2012;32(6):519-524
Objective To determine whether the novel surface-anchored protein SasX promotes aggregation of S.aureus and adherence of S.aureus to human nasal epithelial cells.Methods MRSA ST-239 HS770 sasX gene mutant ( HS770 △sasX) and complement [ HS770 △sasX (pRBsasX) ] were gotten by gene knock-out and complement methods.The aggregation ability of S.aureus was observed through microscope.By adherence assay which was used for detection of the adherence ability of wild type and mutant to human nasal epithelial cells and blocking experiments which detected the ability of the purified recombinant SasX protein in blocking the adherence of S.aureus to human nasal epithelial cells,we investigated the influence SasX on colonization of S.aureus.Results Compared to wild type,HS770△sasX showed a reduction in cell aggregation,while the complement had no difference with wild type in aggregation. HS770 △sasX showed a significant reduction of adherence to human nasal epithelial cells compared to wild type ( P<0.01 ),and the complement showed a very clear increasement of adherence to human nasal epithelial cells compared to wild type(P<0.01 ).Preincubation of nasal epithelial cells with the purified recombinant SasX protein inhibited S.aureus binding significantly.Conclusion SasX had an influence in aggregation of S.aureus and its adherence to human nasal epithelial cells.By acquiring sasX,S.aureus colonized more easily to the susceptible sites of the host,and thus caused infection.
8.Application of Color Doppler Ultrasonography by Bed Side in the Early Diagnosis of Hypoxic-Ischemic Encephalopathy in Full Term Neonates
yi-jin, SU ; lian-fang, DU ; jin, XIA ; min, FANG ; xian-ming, XU ; jian-guo, HONG
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.
9.Application of diffusion weighted echo-planar MR imaging in head and neck lesions
Mu DU ; Jian-Min XU ; Jing-Shan GONG ; Jing-Zhong ZHANG ; Jin ZHU ; Xiao-Mei WANG ;
Chinese Journal of Radiology 2001;0(09):-
Objective to investigate the value of diffusion-weighted echo-planar MR imaging in the diagnosis of head and neck lesions.Methods Fifty-seven patients with 85 head and neck lesions were enrolled in the study,including 22 patients with 22 malignant tumors,13 patients with 13 benign tumors, 13 patients with 17 cystic and liquefactive lesions(including 8 patients with 12 cystic lesions,4 patients with 4 tumor necrosis,1 patients with 1 abcess)and 33 lymph nodes.The lesions were all confirmed by operation and clinical follow up.Echo-planar difffusion-weighted imaging (DWI)was performed with different b values (0,500,and 1,000s?mm~(-2)),and the apparent diffusion coefficients (ADCs)were measured.Results Malignant and benign tumors had different characteristics in DWI with different b values.With the increase of b value,the signal intensity of tumor/spinal cord ratio decreased quickly in DWI in benign tumors,while the signal intensity of tumor/spinal cord ratio remained similar in DWI in malignant tumors.The mean ADC value of'malignant tumors[(0.78?0.24)?10~(-3)mm~2? s~(-1)] was significantly lower than that of benign tumors [(1.48?+0.20)?10~(-3)mm~2?s~(-1)] (t = 8.9,P
10.Purification and Properties of Neutral Protease from Bacillus Subtilis ZC-7
Cong ZHAO ; Min ZHANG ; Jian-Ling WANG ; Lian-Xiang DU ; Xiang-Bin YIN ;
China Biotechnology 2006;0(10):-
Bacillus subtilis ZC-7 was obtained by implantation with N+ ions beam to B.subtilis AS1.398,and compared with the AS1.398 neutral protease,the enzyme activity of ZC-7 neutral protease was about 1 timeshigher in previous research.A neutral protease was purified from the culture of B.Subtilis ZC-7 by the procedures including amoninium sulfate precipitation,ultrafiltration,DEAE-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.By multi-step purification,the ZC-7 neutral protease was purified to 78.5 folds and its yield was 27.7%,at last,the specific activity of ZC-7 neutral protease was up to 4.1?105U/mg.Analysed by SDS-PAGE,the purified protease has shown a molecular mass of about 42kDa.The Km for casein hydrolysis was 3.67?10-3?g/ml and the Vmax was 12.21?g/min.The optimum pH and temperature forhydrolysis of casein were 7.0 and 55℃,respectively.This protease was stable up to 40℃ within the pH range of 6.5 and 8.0.EDTA,isopropanol and alcohol nearly inhibited its activity while some ions such as Ca2+,Mg2+,Fe3+ can improve its activity.In addition,it could resist 1 mol/L H2O2.