1.Fibroblast growth factor 10 inhibits lipopolysaccharide-induced microglial activation
Shulin PAN ; Xiaoxiao HE ; Yingying HU ; Mingchu FANG ; Huai JIANG ; Jian XIAO ; Zhenlang LIN
Chinese Journal of Pathophysiology 2017;33(3):534-538
AIM: To investigate the effects of fibroblast growth factor 10 ( FGF10 ) on lipopolysaccharide ( LPS)-induced microglial activation .METHODS:Mouse BV2 microglial cells were maintained in DMEM in a humidified incubator with 95%/5%( V/V) mixture of air and CO 2 at 37℃.The medium was changed every 1 or 2 d.The cells were digested and passaged every 4 or 5 d.The BV2 microglial cells were first pretreated with FGF 10 (1 mg/L) for 30 min and then stimulated with LPS (500 μg/L).The medium and the cells were collected at different time points .The morphologi-cal changes of microglia were visualized under microscope .To evaluate the microglial activation , the transcription and pro-duction of proinflammatory factor tumor necrosis factor-α( TNF-α) were examined by real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively.RESULTS:The morphology of control BV2 microglia showed circular or oval shape .After exposure to LPS for 24 h, the microglia revealed spindle shaped or multipolar morphology , and the percentage of activated cells was significantly increased compared with control group.Pretreatment with FGF10 successfully inhibited the morphological change from normal to activated shape .LPS sti-mulation for 6 h significantly increased the transcription of TNF-α, while FGF10 pretreatment remarkably reversed the effect.In addition, the production of TNF-αincreased in the presence of LPS stimulation for 24 h compared with control group.Pretreatment with FGF10 suppressed LPS-induced TNF-αexpression.CONCLUSION: Pretreatment with FGF10 inhibits the morphological change from normal to activated shape , and remarkably suppressed the transcription and produc-tion of TNF-α.FGF10 successfully suppresses LPS-induced BV2 microglial activation , indicating that FGF10 is a therapeu-tic agent for the treatment of glia-mediated neuroinflammatory diseases .
2.An analysis of CD3+CD56+ lymphocytes and their subsets in the peripheral blood of patients with chronic hepatitis B.
Peng-jian WENG ; Hao YING ; Ling-zhen HONG ; Wen-hong ZHOU ; Yao-ren HU ; Chen-huai XU
Chinese Journal of Hepatology 2008;16(9):654-656
OBJECTIVESTo investigate CD3+CD56+ lymphocytes and their subsets in the peripheral blood of chronic hepatitis B patients and to explore the relationship between these cells and the pathogenesis of their diseases.
METHODSBlood samples from 53 chronic hepatitis B patients, 17 from HBV asymptomatic carriers (ASC) and 19 from healthy controls (HC) were collected. CD3+CD56+ lymphocytes were detected by flow cytometry (FCM), then the CD3+CD56+ lymphocytes were gathered to analyze their expressions of CD4, CD8, TCR Valpha24, TCRalpha/beta and TCRgamma/delta.
RESULTSThe number of CD3+CD56+ lymphocytes of chronic hepatitis B patients (7.4+/-4.6%) was more than those of ASC (4.5%+/-3.5%) and healthy controls (4.4%+/-3.7%). The expressions of TCR Valpha24 on CD3+CD56+ lymphocytes showed no significant differences among the three groups, but the expression of TCR Valpha24 on CD3-CD56+ lymphocytes of ASC ( 2.8%+/-1.4% ) was much more than that of the HC (1.7%+/-1.0%). For the subsets analysis, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of chronic hepatitis B (61.9%+/-16.8% and 68.1%+/-16.9%) were significantly higher than those of the HC (49.2%+/-15.6% and 56.4%+/-17.9%), while the TCRgamma/delta subsets of chronic hepatitis B and ASC (29.6%+/-15.4% and 30.5%+/-14.8%) were decreased significantly than those of the HC (41.4%+/-19.4%). On the other hand, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of severe chronic hepatitis B (69.0%+/-14.0% and 76.1%+/-12.9%) and CD8 subsets of moderate chronic hepatitis B patients (66.4%+/-14.9%) were significantly higher than those of the mild chronic hepatitis B patients (51.4%+/-16.2% and 62.1%+/-14.6%).
CONCLUSIONThe pathogenesis of chronic hepatitis B may positively relate to the high expression of CD8 on the CD3+CD56+ lymphocytes.
Adult ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Young Adult
3.Establishment of Animal Model for Study of Children with Moderate Blood Lead Levels with Young Rabbits
gao-yan, YU ; chong-huai, YAN ; xiao-gang, YU ; yong, ZUO ; xiang-yu, ZOU ; sheng-hu, WU ; jian, XU ; yan-ping, ZHANG
Journal of Applied Clinical Pediatrics 2006;0(19):-
Objective To establish the animal model for the study of children with moderate blood lead levels in young rabbits,for the study of the ideal therapy for moderate lead poisoning in children.Methods Sixteen 45-day-old male New Zealand rabbits were randomly divided into control and lead-exposed group,8 in each group.Rabbits in the lead-exposed group were treated with 5 mg/(kg?d)lead acetate in their forage for 6 weeks to establish moderate lead poisoning animal model.The blood lead levels(BLLs)were determined by atomic absorption spectrometer(AAS),and the urine lead levels and the lead concentrations of tissue and organ were determined by inductively coupled plasma-mass spectrometry(ICP-MS).Histopathology in tissue and organ was observed under the light microscope.Results The BLLs and the urine lead levels in lead-exposed group step up rapidly in primal weeks,then retained at a steady levels.The BLLs exhibited moderate level BLLs during the lead exposure period.Compared with control group,the body weight gain,testis and hippocampus wet coefficient of the lead-exposed group significantly decreased(P_a
4.Effect and its mechanism of FoxO3a activates FasL on renal tubular epithelial cell apoptosis induced by renal ischemia/reperfusion injury
Jian XU ; Yan WANG ; Huai-Xue JI ; Shu-Qun HU ; Hong-Yan DONG ; Ling REN
The Chinese Journal of Clinical Pharmacology 2014;(10):929-931
Objective To explore the role and mechanism of forkhead box proteinO3a activates Fas ligand on renal tubular epithelial cell ( RTC ) apoptosis induced by renal ischemia /reperfusion ( I/R ) . Methods The model by clamping renal pedicles for 45 minutes follow-ing reperfusion was established.The protein expression of forkhead box proteinO3a and Fas ligand were examined by western blotting.Apoptosis of RTC was assessed by TdT -mediated dUTP nick -end Labeling (TUNEL) method and transmission electron microscopic (TEM).Renal function was assessed by biochemical automatic analyzer .Results The protein expression level of forkhead box proteinO 3a and Fas ligand was increased significantly following renal I /R at 1 h.RTC nucleus was shrinking, crushing followed renal I /R, and a significant increase in the number of TUNEL -positive cells following renal I /R was displayed com-pared with the sham group.The level of blood urea nitrogen(BUN) and serum creatinine ( Scr) was increased significantly compared with the sham group (P <0.05,P <0.01).Conclusion FoxO3a could be acti-vated during renal I /R, and then up -regulated FasL protein expression , facilitated renal tubular epithelial cell apoptosis , in turn, aggravated renal I /R injury in rats.
5.Effects of moderate lead poisoning on the hippocampus tissue of rabbits in juvenile stage.
Gao-yan YU ; Chong-huai YAN ; Xiao-gang YU ; Yong ZUO ; Xiang-yu ZOU ; Sheng-hu WU ; Jian XU ; Xiao-ming SHEN
Chinese Journal of Preventive Medicine 2008;42(3):160-164
OBJECTIVETo understand the effects of moderate lead poisoning on the hippocampus tissue of rabbits in juvenile stage.
METHODSSixteen 45-day-old male New Zealand rabbits were randomly divided into blank group and lead-exposed group,8 for each group. Rabbits in the lead-exposed group were treated with 5 mg x kg(-1) x d(-1) lead acetate in their forage for 6 weeks to establish a moderate lead poisoning animal model. The blood lead levels and the lead contents in the hippocampus were determined by atomic absorption spectrometer and inductively coupled plasma-mass spectrometry respectively. Histopathology and ultra-microstructure in the hippocampus tissue were observed by light microscope and electron microscope. The NR1, NR2A and NR2B protein expressions in the CA1 hippocampal region were analyzed through immunohistochemical method.
RESULTSCompared with those of blank group, the blood lead levels of lead-exposed group were significant increased, (428.63 +/- 9.46) vs (66.38+/-3.93) microg/L (t = 100.08, P<0.01); and lead contents of hippocampus was significantly increased, (44.57+/-2.03) vs (21.20+/-1.53) ng/g, (t = 26.05, P<0.01); the hippocampus wet weight were significant decreased, (0.735 +/-0.012) vs (0.808+/-0.010), (t =12.97, P<0.01); the coefficient of hippocampus wet weight, was (0.458 +/-0.004) vs (0.476+/-0.005), (t =7.87, P<0.01). The significant declines in both the positive rate of NR1 and NR2A in the CA1 hippocampal region for NR1: (37.44 +/- 2.05)% vs (41.81+/-2.50)% (t = 3.82, P<0.01) and for NR2A: 21.97+/-1.08 vs 25.48+/-1.30 (t =5.89, P<0.01) were also observed. With light microscope and electron microscope, the histopathology and ultra-microstructure of neuron and glial cell in the hippocampus tissue were changed.
CONCLUSIONThe impairment of hippocampus of rabbits in juvenile stage with chronic moderate lead poisoning were observed, and the histopathology and N-methyl-D-aspartate receptor protein expressions in the hippocampus tissue were changed.
Animals ; Chronic Disease ; Disease Models, Animal ; Hippocampus ; drug effects ; metabolism ; pathology ; Lead Poisoning ; metabolism ; Male ; Rabbits ; Receptors, N-Methyl-D-Aspartate ; metabolism
6.Epidemiological study on an outbreak caused by E. coli O157:H7 in Jiangsu province.
Hua WANG ; Huai-qi JING ; Hong-wei LI ; Da-xin NI ; Guang-fa ZHAO ; Ling GU ; Jin-chuan YANG ; Zhi-yang SHI ; Guang-zhong LIU ; Xiao-shu HU ; Jian-guo XU
Chinese Journal of Epidemiology 2004;25(11):938-940
OBJECTIVETo carry out epidemiological study on an outbreak caused by E. coli O157:H7 infection in Jiangsu province in 1999.
METHODSEpidemiological, microbiological and moleculebiological methods were used to find out the source, route of transmission and risk factors.
RESULTS95 severe O157:H7 infected patients with acute renal failure in 9 counties and districts of 2 municipalities were reported in Jiangsu province, 1999 while 83 of the patients died with a death rate of 87.37%. Most patients were seen in mid or late June. The ratio of male to female was 1 to 1.44 and 88.42% of the patients were over 50 years old. 38 patients occurred in 2000 with 34 deaths. Major factors contributing to the outbreak would include without drinking tap water, eating leftover food, poor sanitary status in kitchen, not washing hands before meal and after bowl movement. 2 strain of O157:H7 was isolated from severe patients and 3 from diarrhea cases. Carrier rate among animals was up to 9.62% and 99.41% of the strains carried toxic gene. Strains isolated from feces of patients and animals belonged to the same colonies.
CONCLUSIONThis outbreak was severe which caused by O157:H7 and was first seen in China, which was closely related to the high carrier rate of O157:H7 in animals and to the positive rate of high toxic gene of the strains. There were various routes of transmission and the main factors of infection would include poor personal health habits and poor sanitation of the household.
Acute Kidney Injury ; epidemiology ; etiology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Bacterial ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; microbiology ; Disease Outbreaks ; Escherichia coli Infections ; complications ; epidemiology ; Escherichia coli O157 ; isolation & purification ; Escherichia coli Proteins ; immunology ; Female ; Hemolysin Proteins ; immunology ; Humans ; Infant ; Male ; Middle Aged ; Seroepidemiologic Studies
7.Overexpression of COX-2 and its clinical significance in non-small cell lung cancer.
Feng TIAN ; Tian-you WANG ; Min GONG ; Xiao-mei LU ; Jian HU ; Jing WANG ; Chang-huai ZHANG
Chinese Journal of Surgery 2003;41(6):407-410
OBJECTIVETo study the relations among COX-2 expression in non-small cell lung cancer (NSCLC), its clinical characteristics and brognosis.
METHODSImmunostaining was performed with COX-2 antibody to the surgically resected tissue samples from 79 patients with NSCLC. Vessel epithelium cell COX-2 expression was taken as positive control.
RESULTSThe positive rate of adenocarcinoma and squamous cell carcinoma was 85% and 57%, respectively (P = 0.013). COX-2 expression was associated with the extent of adenocarcinoma differentiation, tumor size, and TNM period, but not with the extent of squamous cell carcinoma differentiation. In the COX-2 positive group, the 5-year survival rate and a median survival time were 27.1% and 53 months; however in the negative group they were 52.0% and 61 months (P = 0.029).
CONCLUSIONInvasive development of NSCLC is related to inceased expression of COX-2. COX-2 overexpression may be one of the risky factors for the prognosis of NSCLC.
Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; enzymology ; mortality ; pathology ; Cyclooxygenase 2 ; Female ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Lung Neoplasms ; enzymology ; mortality ; pathology ; Male ; Membrane Proteins ; Middle Aged ; Prognosis ; Prostaglandin-Endoperoxide Synthases ; analysis ; Survival Rate
8.The roles of c-Jun and CBP in the inhibitory effect of quercetin on prostate cancer cells.
Hui-qing YUAN ; Huai-fang GUO ; Mei-lan HE ; Feng KONG ; Xiao-Yan HU ; An-li JIANG ; Xia XU ; Jian-ye ZHANG ; Y F Young CHARLES
Acta Pharmaceutica Sinica 2006;41(9):819-824
AIMTo further uncover the possible mechanism of quercetin-mediated inhibitory effect on prostate cancer cells.
METHODSThe cell extracts treated with quercetin or without treatment were used for checking protein expression levels of c-Jun and cAMP response element binding protein (CREB)-binding protein (CBP) by Western blotting assay. Regulatory effects of c-Jun and CBP on the function of androgen receptor (AR) were examined by cotransfection experiment. Finally, a physical interaction of c-Jun and the AR was investigated by coimmunoprecipitation.
RESULTSQuercetin dramatically induced the protein expression of c-Jun which in turn inhibited the AR function. Meanwhile, quercetin had no detectable effect on CBP expression, and the results of transient transfection demonstrated that the ectopic CBP stimulated the transcriptional activity of AR, whereas CBP-mediated stimulation could be attenuated by quercetin. Furthermore, physical interaction of c-Jun and the AR was confirmed by coimmunoprecipitation result.
CONCLUSIONOverexpression of c-Jun induced by quercetin had inhibitory effect on the function of AR protein, and increased CBP expression did not reverse the inhibition by quercetin. Together, quercetin-mediated inhibition on the AR function might be not by competition with limited amount of CBP in the cell, but through a direct association of c-Jun and the AR.
Antineoplastic Agents, Phytogenic ; pharmacology ; CREB-Binding Protein ; genetics ; metabolism ; physiology ; Cell Line, Tumor ; Humans ; Immunoprecipitation ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Protein Binding ; drug effects ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; physiology ; Quercetin ; pharmacology ; Receptors, Androgen ; genetics ; physiology ; Transfection
9.Arsenic trioxide inhibits P-glycoprotein expression in multidrug-resistant human leukemia K562/ADM cell line that overexpresses mdr-1 gene and enhances their chemotherapeutic sensitivity.
Hu-lai WEI ; Xiao-jian YAO ; Yu-ning LI ; Pei WANG ; Huai-shun ZHAO ; De-cheng BAI ; Xiao PENG ; Lan-fang MA
Chinese Journal of Hematology 2003;24(1):28-31
OBJECTIVETo investigate the effects of arsenic trioxide (As(2)O(3)) on the apoptosis and P-glyco-protein (P-gp) expression of multidrug-resistant human leukemia K562/ADM cells, and the combined effects of As(2)O(3) with conventional chemotherapeutic agents.
METHODSMultidrug-resistant human leukemia cell line K562/ADM that overexpresses mdr-1 gene was used as the target cells. The cell proliferating activity was assessed with a MTT assay. Cell morphology was examined by light microscopy, confocal microscopy and electron-microscopy. P-gp expression, cell-cycle status were determined by flow cytometry.
RESULTSK562/ADM cells were highly resistant to adriamycin, and cross-resistant to daunorubicin and etoposide. As(2)O(3) at concentrations of 0.5 to 20 micromol/L inhibited the proliferation of K562/ADM cells, and K562/ADM cells were more sensitive to As(2)O(3) than their parent K562 cells did. As(2)O(3) induced marked apoptosis of K562/ADM cells showed by typical apoptotic morphological changes and the appearance of high sub-G(1) cell population. As(2)O(3) significantly inhibited the P-gp expression in K562/ADM cells, and exerted a synergistic effect on the enhancement of the cell sensitivity to adriamycin, daunorubicin and etoposide.
CONCLUSIONAs(2)O(3) induces growth-inhibition and apoptosis of multidrug-resistant K562/ADM cells, and augments synergistically the sensitivity of the cells to conventional chemotherapeutic agents via down-regulation of P-gp expression.
ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; drug effects ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Daunorubicin ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drug Synergism ; Etoposide ; pharmacology ; Humans ; K562 Cells ; Oxides ; pharmacology
10.Analysis of prosthetic replacement in treatment of femoral neck fracture on the hemiplegia side in the elderly.
Ming-li FENG ; Hui-liang SHEN ; Huai-jian HU ; Yi-min YONG ; Li CAO ; Wei WANG
Chinese Journal of Traumatology 2004;7(3):138-142
OBJECTIVETo study the effect of prosthetic replacement in treatment of femoral neck fractures on the hemiplegia side in the elderly.
METHODSFrom May 1990 to May 2000, 189 elderly patients with femoral neck fractures were treated with prosthetic replacement in my hospital. Twenty-nine hemiplegia patients, who suffered from stroke previously, had Garden type III and type IV femoral neck fractures on the hemiplegia side. Thirty non-hemiplegia patients were chosen randomly. The two groups were followed-up for 27-98 months (average: 59 months). The age, hospitalization days, operating time, blood loss, blood transfusion, complications during perioperative period and long-term complications were compared between the two groups and the results of femoral head replacement and total hip replacement in the hemiplegia group were also compared.
RESULTSAll the patients of the two groups survived the perioperative period. No significant difference was found in the age, hospitalization days, operation time, blood loss and blood transfusion and long-term complications between the two groups (P>0.05). However there was significant difference in complications during perioperative period between the two groups (P<0.05). Five patients died in the hemiplegia group with the mortality of 17.2% and two died in the non-hemiplegia group with the mortality of 6.7% 11 months to 5 years after operation. There was significant difference in long-term complications between the femoral head replacement and the total hip replacement in the hemiplegia group (P<0.05). The result of the total hip replacement was better than that of the femoral head replacement.
CONCLUSIONSProsthetic replacement is a reliable method in treatment of Garden type III and type IV femoral neck fractures on the hemiplegia side in the elderly, and patients are safe during perioperative period. More complications during perioperative period occur in the hemiplegia group, and long-term complications are insignificantly different between the two groups. The mortality rate is higher in the hemiplegia group than in the non-hemiplegia group within 5 years after operation. Since the result of the total hip replacement is better than that of the femoral head replacement, total hip replacement should be chosen firstly to treat Garden type III and type IV femoral neck fractures on the hemiplegia side in the elderly if the muscular strength of the hip is beyond IV degree.
Aged ; Arthroplasty, Replacement, Hip ; Female ; Femoral Neck Fractures ; etiology ; surgery ; Hemiplegia ; complications ; Humans ; Male