2.Effect of Modified Qinghao Biejia Decoction on Th17 Cells and Renal Pathology of MRL/lpr Mice with Lupus Nephritis
Ning LIN ; Jiaxi ZHONG ; Bin QIU ; Jieying FENG ; Xia SUN
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(5):776-779,785
Objective To observe the effect of modified Qinghao Biejia Decoction ( QBD) on Th17 cells and renal pathology of MRL/lpr mice with spontaneous systemic lupus erythematosus. Methods Thirty-two female MRL/lpr mice aged 8 to 10 weeks were divided into 4 groups: model group, Chinese medicine group, prednisone group, and combination group, 8 mice in each group. Eight female C57BL/6 mice aged 8 to 10 weeks served as normal control. Mice in Chinese medicine group were given concentrated solution of modified QBD (19.25 g·kg-1·d-1), mice in the prednisone group were given water solution of prednisone acetate (8.75 mg·kg-1·d-1) , mice in the combination group were given the above two kinds of medicine, and mice in the model group and normal control group were given physiological saline. After medication for 7 weeks, spleens and kidneys in all of the groups were taken out for the experiment. Th17 cells in splenic mononuclear cell suspension were detected by flow cytometry, the pathological changes of renal tissue were observed under light microscope, and activity index (AI) of renal tissue in lupus nephritis mice was scored. Results The proportion of Th17 cells in the model group was significantly higher than that of normal control group ( P<0.05) . The proportion of Th17 cells in Chinese medicine group and combination group was lower than that of the model group ( P<0.05) , and prednisone group had higher proportion of Th17 cells than Chinese medicine group ( P<0.05) . Compared with the model group, pathological changes of renal tissue were relieved, and AI scores were decreased in Chinese medicine group, the prednisone group and the combination group ( P<0.05) . Except for the normal control group , AI scores in all groups were positively correlated with the proportion of Th17 cells ( r=0.77, P<0.01) . Conclusion Modified QBD can inhibit the expression of Th17 cells and improve the pathological changes of MRL/lpr mice with lupus nephritis.
3.Pathological morphology of vascular endothelial injury and arteriosclerosis caused by high fluoride and the effects of selenium
Jian-chao, BIAN ; Xiao-xia, YANG ; Xin-ying, LIN ; Qiu-li, ZHU ; Ting, FAN
Chinese Journal of Endemiology 2010;29(6):593-598
Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P < 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P < 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P < 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P < 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P < 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P < 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.
4.Detecting multiple lysosomal enzymes in dried blood spots by tandem mass spectrometry
Xia ZHAN ; Xuefan GU ; Na LIN ; Jun YE ; Wenjuan QIU ; Lianshu HAN ; Huiwen ZHANG ; Lili LIANG
Chinese Journal of Laboratory Medicine 2016;39(10):761-765
Objective Establish a method for measuring the activities of Galactocerebrosidase (GALC), α-Glucosidase(GAA), α-Galactosidase (GLA) and α-L-Iduronidase (IDUA) in dried blood spots specimen by tandem mass spectrometry ( MS/MS ).Methods A total of 2175 dried blood spot samples forinborn errors of metabolism in neonatalscreening center of Shanghai Xinhua hospital were collected in July and November, 2013.And twenty dried blood spot samples from patients withlysosomalstorage disorders( LSDs) of Shanghai Xin Hua Hospital were collected from September 2012 to January 2014.The extraction of DBS was incubated with enzyme substrates and internal standards.After liquid-liquid and solid-phase extraction, the extraction solution was dried under nitrogen and reconstituted.Then enzyme reaction products and internal standards were analyzed by MS/MS.Linearity, precision, accuracy and the limit of detection were evaluated.2175 dried blood spot samples were detected to establish the normal reference range for the activities of four enzymes according to 0.5th to 99.5th percentiles.20 specimens from patients withLSDs were detected to verify the reference range inclinical judgment.Results The intraassay and interassay precisions ranged from 1.7%to 11.8%, and the intraassay and interassay accuracies ranged from 85%to 115%.The linear coefficients for measured concentration of enzyme products/internal standards and theoretical concentration were 0.997-0.999.The limits of detection forGALC, GAA, GLA and GLA were 0.03 μmol/(L· h), 0.09 μmol/(L· h), 0.12 μmol/(L· h) and 0.16 μmol/(L· h) .The normal reference values for GALC, GAA, GLA and GLAwere 0.51-8.51μmol( L· h) ,1.99-22.22μmol/( L· h),1.68-41.59 μmol/(L· h) and 2.36-19.21 μmol/(L· h).The enzymes of 20 patients with LSDs were remarkably decreased compared to the normal range.The Krabbe, Pompe, Fabry, MPSⅠpatients can be effectively detected by this MS/MS method.Conclusions A MS/MS method for measuring GALC, GAA, GLA and IDUA enzyme activities in DBShas been established.
5.Human umbilical vein vascular endothelial cell injury induced by fluoride in vitro
Jian-chao, BIAN ; Xin-ying, LIN ; Xiao-xia, YANG ; Xiao-dong, HOU ; Ting, FAN ; Qiu-li, ZHU
Chinese Journal of Endemiology 2011;30(2):142-147
Objective To study the effect of different concentrations of fluoride on cultured human umbilical vein vascular endothelial cells(HUVEC). Methods Different doses of sodium fluoride (NaF) were added to HUVEC culture medium, fluoride concentrations were 0(control), 100,400,700,1000,2000 μmol/L, respectively,6 re-set hole in each group. After continuous culture for 48 h, cells and culture medium were collected. Cell morphology was studied by Wright-Giemsa staining; cells apoptosis was determined by acridine orange fluorescence staining; cell activity was measured by methyl thiazolyl tetrazolium (MTT) assay; superoxide dismutase (SOD),glutathione peroxidase(GSH-Px) activity, malonaldehyde(MDA) content, induced nitricoxide synthase(iNOS), and endothelia nitricoxide synthase(eNOS) activity in cell culture medium were determined by spectrophotometry; cell iNOS mRNA and eNOS mRNA expression were detected by RT-PCR; intercellular adhesion molecule-1 (ICAM-1)and vascular cell adhesion molecule-1 (VCAM-1) levels were detected by double antibody sandwich ELISA method.Results With increased dose of fluoride, HUVEC cells decreased, the structure changed. In 400 - 2000 μmol/L group, the SOD activity[(6.627 ± 0.213), (6.668 ± 0.152), (5.935 ± 0.122), (4.755 ± 0.182)kU/L] was lower than those of the control group[(7.457 ± 0.398)kU/L, P < 0.05 or < 0.01], GSH-Px activity[(481.284 ± 43.785),(492.223 ± 16.474), (382.762 ± 25.167), (293.687 ± 24.881 )kU/L] was also lower than those of the control group [(585.078 ± 47.323)kU/L, P < 0.05 or < 0.01], MDA level[(0.609 ± 0.011 ), (0.646 ± 0.016), (0.852 ± 0.013),(1.188 ± 0.045)nmol/L] was higher than those of the control group[(0.512 ± 0.027)nmol/L, P < 0.05 or < 0.01];iNOS activity[(3.604 ± 0.115), (3.615 ± 0.075), (3.848 ± 0.103), (4.275 ± 0.079)kU/L] also was higher than those of the control group[(2.798 ± 0. 136)kU/L, all P < 0.01], iNOS mRNA expression increased, eNOS activity [(5.539 ± 0.079), (5.503 ± 0.064), (5.226 ± 0.142), (4.809 ± 0. 107)kU/L] decreased compared to those of control group[(5.996 ± 0.155)kU/L, P < 0.05 or < 0.01], eNOS mRNA expression decreased; ICAM-1 levels [(0.852 ± 0. 102), (0.886 ± 0.061 ), (0.961 ± 0.158), (1.418 ± 0. 167)μg/L] increased compared to those of the control group[(0.687 ± 0.046)μg/L, P < 0.05 or < 0.01], VCAM-1 levels[(2.719 ± 0.197), (2.946 ± 0.167),(3.173 ± 0.225 ), (3.613 ± 0. 153 ) μg/L] was higher than those of the control group [(2.375 ± 0.067 ) μg/L, all P <0.01]. Conclusions High concentrations of fluoride reduce the activity of antioxidant enzymes, which leads to metabolic disorders of nitric oxide and abnormal cytokines expression, thereby inhibiting vascular endothelial cell growth, structural change and induced apoptosis. This is an important factor in high fluoride-induced vascular endothelial injury.
6.Recurrent low frequency sensorineural deafness.
Ying LIN ; Jin Ling WANG ; Fei SUN ; Jin Jin SHEN ; Zhao Xia WANG ; Jian Hua QIU ; Ding Jun ZHA
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2018;32(6):474-476
Low frequency sensorineural deafness is a common subtype of idiopathic sudden deafness. Certain patients suffered recurrent attacks without vertigo, much alike Meniere's disease. Few of them developed into definite Meniere's disease during long-term follow-up in many clinical studies. Although the pathophysiology of recurrent low frequency deafness is yet unknown, the desease is considered associated with early state of endolymphatic hydrops or migraine. Otologists shall be aware of its clinical course and careful explanation is necessary at time of initial informed consent.
Endolymphatic Hydrops
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complications
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Hearing Loss, Sensorineural
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complications
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diagnosis
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Hearing Loss, Sudden
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Humans
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Meniere Disease
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complications
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Vertigo
7.Simultaneous kidney-pancreatic transplantation with portal venous and enteric drainage:a report of 6 cases
Lin ZHONG ; Zhihai PENG ; Zhengjun QIU ; Qiang XIA ; Guoqing CHEN ; Zhecheng ZHU ; Junming XU ; Jun LIU ; Shuyun WANG
Chinese Journal of General Surgery 2001;0(07):-
Objective To evaluate clinical effect of simultaneous kidney-pancreatic transplantation(SKPT) with portal venous and enteric drainage. Method Between June 2001 and June 2004, six insulin-dependent diabetes mellitus(IDDM) and renal failure patients underwent this procedure. The venous drainage of the graft was established between donor′s portal vein and recipient′s superior mesenteric vein. The exocrine secretion was drained into proximal jejunum via side-to-side anastomosis between donor′s duodenum and recipient′s proximal jejunum. Steroids, mycophenolate mofetil, tacrolimus and Zenapax were used as immunosuppressants. Results Procedures were successful in all 6 cases. Excellent renal function and euglycemia were achieved in 4 cases. Follow-up of 4-34 months on the 4 survivers found excellent kidney-pancreatic function without any rejection episode. Two patients died perioperatively due to sepsis secondary to pancreatic leakage and drug toxicosis of excessive FK506. Conclusion Our preliminary experience suggests that simultaneous pancreas-kidney transplantation with enteric and portal drainage is reliable procedure for the treatment of IDDM with renal failure.
8.Induced-division of neurons derived from neural stem cells.
Qiu-Xia LIN ; Hai-Ping QUE ; Shuang-Hong LU ; Shao-Jun LIU
Acta Physiologica Sinica 2004;56(2):130-136
In order to explore if mature neurons derived from neural stem cells have the potentiality to divide, we utilized the chemical digestion method to disperse the adult rat brain tissue into single cells, and culture them in serum-free medium. After being cultured for about eight days in vitro, the neural stem cells were induced to differentiate into neurons. The neurons were further induced to divide. Utilizing the method of serial photograph and NF-160 immunocytochemistry, the processes of division of some neurons were recorded. At the same time, PCNA+NF-160 (or Chat, GABA, GAD) double label were used to investigate if the dividing-neurons were mature ones. After the neural stem cells were induced to differentiate in vitro for eight days, they possessed the shape and character of mature neurons. The differentiated neuron had a big nucleus and one or two distinct nucleolus in the nuclear. Within the perikaryon,there were a large amount of dense and Nissl body-like structure. Several long processes emerged from various locations of the cell body. Then, EGF and bFGF were added into the medium to induce division. After two days of induced-division, neuron-like cells were observed to divide; moreover, the number of neuron-like cells in the region increased continually. Immunocytochemistry demonstrated these cells were NF-160-positive. Serial photographs of dividing-process of neuron-like cells were obtained and their daughter cells were also NF-160-positive. After PCNA+NF-160 (or Chat, GABA, GAD) double label, some cells showed brown cell plasma and black nucleus. The above-mentioned results indicate that neurons, which were previously thought to be end-differentiated, can be re-called into cell cycle under appropriate conditions. Mature neurons still have the potential to divide, proliferate and self-renew.
Animals
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Brain
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cytology
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Cell Differentiation
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Cell Division
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Cell Separation
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Cells, Cultured
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Epidermal Growth Factor
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pharmacology
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Fibroblast Growth Factor 2
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pharmacology
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Neurons
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cytology
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Photography
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methods
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Proliferating Cell Nuclear Antigen
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pharmacology
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Rats
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Rats, Wistar
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Stem Cells
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cytology
9.Autologous hematopoietic stem cell transplantation in the treatment of patients with poor-prognosis non-Hodgkin iymphoma
Bo CHEN ; Zhiguo WANG ; Liang XIAO ; Hao ZHANG ; Yanling FAN ; Guoqiong XIA ; Lin QIU ; Zhaomin ZHAN ; Bolong ZHANG ; Jun MA
Journal of Leukemia & Lymphoma 2009;18(10):606-608
Objective To observe the therapeutic effectiveness and safety of autologous peripheral blood stem cell transplantation (APBSCT) for poor-risk non-Hodgkin lymphoma (NHL). Methods Ten patients with poor-prognosis NHL were enrolled from October 2003 to October 2008 in our institute. Ten patients were treated by APBSCT with CY+TBI conditioning regimen (Two patients of them were treated by Double-APBSCT with MEC conditioning regimen). Results Hematopoietic reconstitution was observed in all patients.The time of neutrophil count ≥0.5×109L and platelet≥20×109/L were at day 10 (range: 7-14) and day 16 (range: 10-37), respectively. All patients achieved complete remission (CR) after transplantation. Severe toxicity and transplant related mortality were not observed. After a median follow-up of 24 (10-84) months,seven cases were in event-free survival and three cases relapsed. One of three relapsed patients died from progression of disease, the other was still alive after treatment. Conclusion APBSCT in the treatment of patients with poor-prognosis NHL is a safe, convenient and efficient treatment.
10.Purification and part of physico-chemical characterization as well as biological activity for immuno-globulin ribonucleic acid of anti-rabies
Lin YANG ; Suzhi YAN ; Jinyan ZHANG ; Chunhong LIU ; Ye QIU ; Tong LI ; Yingxin XU ; Tianyao XIA ; Su YAN
Chinese Journal of Microbiology and Immunology 2010;30(2):140-143
Objective To explore the preparation of specific immune RMA(iRNA) on anti-rabies and further study immunotherapy of rabies virus exposure. Methods Horses were immunized with the rabi-es virus and their livers were isolated from the horse of antiserum, from which total RNA was extracted and purified by sodium lauryisulfonate, phenol, chloroform, ethyiene glycol monomethyl ether, cetyltrimethyam-moniumbromide and alcohol. Results Pure preparation physico-chemical characterization was analyzed, and it's weight was 0.15% of weight of liver. The RNA contained 2.86% DNA and 1.16% protein. The iRNA with a maximum UV absorbance at 258 nm and A_(258/280) about 2.0. The test of RNA was positive, which had a relative molecular mas of 13.7×10~3 by high performance liquid chromatography(HPLC), and its hy-perchromic effect was 50.67%. The vesults of biological activity was showed that the rate of leucocyte adher-ence inhibition(LAI) was 41.73%, The protective rate was 50% and prolonging the life was 31.62%. Conclusion The results obtained with the practical value were identical and provide a basis on medicines of anti-rabies.