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 .

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

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

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

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

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

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

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

OBJECTIVETo explore the effect of small interfering RNA(siRNA) on silence of mdr1 gene and reversal of apoptosis resistance in multidrug-resistant (MDR) human leukemia K562/ADM cell.

METHODSHuman MDR leukemia cell line K562/ADM was used as the target cells. Two siRNAs (mdr1 siRNA-1 and mdr1 siRNA-2) targeted mdr1 gene were chemically synthesized and transfected into K562/ADM cells with liposome. Expression of mdr1 mRNA was determined by real-time PCR, P-glycoprotein (P-gp) expression and caspase-3 activity were measured with flow cytometry (FCM), and the cell apoptosis was observed by optical and electronic microscopy for morphology and Annexin V/PI staining.

RESULTSThe mdr1 siRNA-1 and mdr1 siRNA-2 could markedly down-regulate the expression of mdr1 gene in K562/ADM cells, the expression of mdr1 mRNA decreased by 91.2% and 82.0% , and the P-gp by 74.1% and 84.4%, respectively. The caspase-3 activity was markedly enhanced, and the active caspase-3 in K562/ADM cells increased by about 40% compared to liposome alone and non-silencing controls. the sensitivity of K562/ADM cells to adriamycin-induced apoptosis was significantly augmented, the apoptotic rate of the cells treated with siRNA plus adriamycin increased by about 60% compared to adriamycin alone.

CONCLUSIONsiRNAs silence the expression of mdr1/P-gp to overcome the P-gp-mediated apoptosis resistance in drug-resistant K562/ADM cells.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Caspase 3 ; metabolism ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

This study was aimed to investigate the killing activity of cytokine-induced killer (CIK) cells after being incubated with autologous tumor cell lysate-pulsed dendritic cells (DC) and to evaluate the clinical efficacy and side effect of autologous tumor cell lysate-loaded DC in combination with CIK on relapsed or refractory non-Hodgkin's lymphoma (NHL). Peripheral blood mononuclear cells (PBMNC) were isolated from 9 patients with NHL, and cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) to produce DC. The DC were pulsed with autologous tumor cell lysate. T lymphocytes from PBMNC were cultured with interferon-gamma (IFN-gamma), IL-2, CD3-moAb, and IL-1alpha to prepare CIK. After receiving the immunotherapy of DC and CIK, immunologic and clinical responses were evaluated. The results showed that the AgNOR, CD3(+)CD8(+) and CD3(+)CD56(+) ratio were markedly improved after the immunotherapy (p < 0.01); IFN-gamma and IL-12 levels in supernatant of DC-CIK group were higher than that in CIK group (p < 0.01); Tumor size were significantly decreased after the immunotherapy (p < 0.05). Except transient fever and chill, no remarkable adverse event happened during or after the treatment. It is concluded that the autologous tumor cell lysate-pulsed DC in combination with CIK show ability to specifically kill the lymphoma cells, obviously increases the IS value of Ag-NOR in peripheral lymphocytes, secretes cytokines higher than CIK cells alone. This combination displays the short-term satisfied efficacy on NHL through inducing specific antitumor immunity, and can be used as an effective adjuvant measure for the routine therapy of NHL.
Adult ; Aged ; Cytokine-Induced Killer Cells ; immunology ; Dendritic Cells ; immunology ; Female ; Humans ; Immunotherapy ; Immunotherapy, Adoptive ; Lymphoma, Non-Hodgkin ; immunology ; therapy ; Male ; Middle Aged ; Recurrence ; Treatment Outcome