Objective To establish a flow cytometry-based assay for the detection of monocyte-me-diated antibody-dependent cell-mediated cytotoxicity ( ADCC ) .Methods P815 cells double stained with PKH26 and carboxyfluorescein succinimidyl ester ( CFSE ) were used as target cells and coated with P 815 specific antibodies to form antigen-antibody complexes .The peripheral blood mononuclear cells were isolated as effector cells and co-cultured with the antigen-antibody complexes .The CD3-CD14+PKH26+CFSE-cell population were gated by flow cytometry .Optimized effector/target cell ratio and incubation time for killing assay were identified .Monocyte-mediated ADCC in 23 patients with chronic HCV infection and 22 healthy subjects were analyzed .Results The monocyte-mediated ADCC could be evaluated through analyzing the CD3-CD14+PKH26+CFSE-cells with flow cytometry .The optimized effector/target cell ratio was 10 ∶1 and the optimized time for incubation was 4 h.Monocyte-mediated ADCC was inhibited in patients with chronic HCV infection as compared with healthy subjects (P=0.009).Conclusion A flow cytometry-based assay for the detection of monocyte-mediated ADCC was established , which could be used as a fast , sensitive and safety method for the evaluation of monocyte-mediated ADCC during viral infections and the research and de-velopment of drugs .

Benzofurans ; analysis ; chemistry ; Dibenzofurans, Polychlorinated ; Dioxins ; analysis ; chemistry ; Humans ; Occupational Exposure ; prevention & control

Objective To investigate the organophosphorus pesticides residues in vegetables in Ji'nan,China.Methods From 2005 to 2007,the residues of ten kinds of organaphosphorous pesticides,including methamidophos,parathion-ethyl, parathionm-ethyl,dichlorvos,phorate,monocrotophos,omethoate,dimethoate,chlorpyrifos,malathion were determined in 8 kinds of vegetables by capillary gas chromatography.Results In the 597 vegetables samples,the total detection rate of organaphosphorous pesticides was 32.66%,in 25.63% of the samples,the contents of organaphosphorous pesticides residues exceeded the national standard limits.In leek,the detection rate was 65.56% and the exceeded limit rate was 52.22%.In addition, more than one kind of pesticide could be detected in one kind of vegetable.The concentration of methamidophos,the most serious pollutant,could reach 114.013 mg/kg.Omethoate could also be detected at 11.807 mg/kg.In this investigation,a downtrend was seen in the rate of exceeding standard limits and the difference in seasons in the detection rate of pesticides residues was also seen. Conclusion The detection rate of high toxic organaphosphorous pesticides in vegetables sampled in this study is higher,it indicates that the situation of pesticides abuse is serious in this area.

Acute Kidney Injury ; therapy ; Child ; Hemofiltration ; methods ; Humans

OBJECTIVETo detect angiotensin II (ANG II) in the maternal blood, umbilical cord blood, and at the maternal-fetal interface in women with pregnancy-induced hypertension (PIH) and explore the etiology of PIH and pathophysiologic mechanism of intrauterine growth restriction (IUGR).

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of ANG II in the maternal blood, cord blood, and maternal and fetal placental tissues in 30 women with PIH and 30 with normal pregnancy, and the results were analyzed with independent-sample t test and Pearson correlation analysis.

RESULTSANG II level in the maternal placental tissue homogenate showed no significant difference between women with PIH and normal pregnant women (8.51+/- 4.01 vs 7.76+/-3.47 pg/ml, P>0.05), but women with PIH had significantly higher ANG II in the fetal placental tissue (11.82+/-3.92 vs 9.64+/-2.63 pg/ml, P<0.05). ANG II level was significantly higher in the maternal blood of women with PIH than in normal pregnant women (46.44+/-8.48 vs 32.43+/-5.87 pg/ml, P<0.001), but similar in the cord blood (68.83+/-8.68 vs 72.47+/-8.51 pg/ml; P>0.05). A positive correlation was indicated between the cord blood and maternal peripheral blood ANG II levels in women with PIH (r=0.7379, P<0.05).

CONCLUSIONANG II in the fetal placental tissue is elevated, and the cord blood and maternal peripheral blood ANG II levels are positively correlated in women with PIH.

Angiotensin II ; blood ; isolation & purification ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Female ; Fetal Blood ; chemistry ; Fetal Growth Retardation ; blood ; Humans ; Hypertension, Pregnancy-Induced ; blood ; Placenta ; chemistry ; Pregnancy

OBJECTIVETo study the relationship between gene p53 codon 72 polymorphism and pathological scar formation occurrence after caesarean section.

METHODSThe method of molecular beacon with real-time PCR was applied to detect gene polymorphism of p53 codon 72 in blood samples taken from 303 pregnant women (within a week after caesarea section). The clinical visits were taken 3 times for 12th to 18th months to ascertain clinical formation of pathological scar and its relationship to genotype of p53. The chi-square method was used to analyze the relationship of p53 gene polymorphism and abnormal scar formation occurrence by statistical software SPSS 13.0.

RESULTSTotal of 303 pregnant women were assayed. 30 patients were found with pathological scar by clinical visit in the total 303 pregnant women. The genotype frequencies of total three types (C/C, C/G and G/G) of p53 gene codon 72 in patients with pathological scar are significantly different from that of normal pregnant woman. The frequency of C/C genotype in patients are higher than that of normal pregnant women (P < 0.01). The frequency of C/C genotype in these patients with pathological scar is higher (46.7%, 14/30) than C/G (33.0%, 10/30, P < 0.01) or G/G (20%, 6/30) genotype (P < 0.01). The C allele frequency in the patients is 63.7%. It is also higher than G allele (36.7%, P < 0.01). The OR value is 2.30. Therefore the C allele of p53 gene codon 72 is a risk factor for pathological scar.

CONCLUSIONSThere was a certain relationship between p53 gene codon 72 C allele and pathological scar formation after caesarean section.

Alleles ; Cesarean Section ; Cicatrix ; genetics ; Codon ; Female ; Gene Frequency ; Genes, p53 ; Genotype ; Humans ; Polymorphism, Genetic ; Pregnancy ; Risk Factors

Objective To study the clinical features and prevention means of the infection caused by the highly dathogenic avian influenza A(H_5N_1).Method The clinical data which were confirmed to be an H_5N_1 infected case were analyzed retrospectively.Results The patient,16-year-old male,had no unambi guous history of direct contact with diseased or dead poultry before the onset of the disease.After the onset of the disease,chest X-ray showed flake shadow of the right lower lung quickly spread to the whole lung,associated with mediastinum,subcutaneous emphysema,acute respiratory distress syndrome(ARDS)was occurred on the fifth day.Mechanical ventilation is the primary measure in the comprehensive treatment.On the sixth day,Chinese Center for Disease Control and Prevention detected the pharyngeal specimen of the patient by RT - PCR,Real- time PCR method and suggested positive for the A/H5/N1 virus nucleic acid andis01ated the avian flu(H_5N_1) virus.The disease course was 10 days from onset of illness to death.Conclusions Human infection by the highly pathogenic avian influenza A(H_5N_1)is a deadly infectious disease.If the lesions is widespread and associated with ARDS,prognosis is poor.

OBJECTIVETo explore the clinical and physical factors that might give rise to radiation-induced esophagitis in three-dimensional conformal radiotherapy for non-small cell lung cancer.

METHODSTo collect the clinical and physical records and follow-up information of 106 NSCLC patients without undergoing surgery in our hospital. χ(2) test, linear tendency test and analysis of variance were employed to analyze the relationship between occurrence of radiation-induced esophagitis and clinical and physical treatment. Logistic analysis was also used for multivariate analysis.

RESULTSAmong the 47 cases of radiation-induced esophagitis, 31 cases were of grade I, 11 of grade II, 5 of grade III, and with a total occurrence rate of 44.3% (47/106). Radiation-induced esophagitis was correlated with Karnofsky scores, radiation sensitization and tumor location (χ(2) = 11.30, 8.45, 7.67, P < 0.05). Radiation-induced esophagitis was correlated with the length of irradiated esophagus and average dose of irradiated esophagus (F = 20.82, 83.08, P < 0.001). With the increase of the irradiated volume percentage from V20, V30, V40 up to V50, the occurrence rate of radiation-induced esophagitis was also increased, almost with a linear trend (P < 0.05). Application of all the above factors to logistic model indicated that radiation sensitization,length of irradiated esophagus, average dose and V50 were all statistically significant foactors in the occurrence of radiation-induced esophagitis (OR = 0.321, 2.850, 7.307 and 8.558, P < 0.05).

CONCLUSIONSRadiation sensitization,length of irradiated esophagus, average dose of irradiated esophagus and V50 are independent factors in the occurrence of radiation-induced esophagitis. V50 is of greater importance in the judgement of occurrence of radiation-induced esophagitis.

Aged ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Esophagitis ; etiology ; Esophagus ; pathology ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Multivariate Analysis ; Radiation Injuries ; etiology ; Radiation Tolerance ; Radiotherapy Dosage ; Radiotherapy, Conformal ; adverse effects ; methods ; Retrospective Studies

OBJECTIVETo investigate the molecule mechanism of the anti-fibrotic effects of Chinese herbal drugs (Qidan granules) in rats.

METHODSThe male rats were randomly divided into four experimental groups: normal group, model group, Qidan group, tetrandrine group. Every group had 10 rats. Normal group were treated with physiologic saline while others were treated with silicon dioxide (50 mg/rat) by intratracheal instillation to induce silicosis. On 30th day Qidan group and Tetrandrine group were treated with Qidan granules (3125 mg/kg) or treated with tetrandrine (22 mg/kg) respectively. All the rats were scarified after 5 months. Lung/body coefficient was measured. Content of hydroxyproline was measured by alkaline hydrolysis. The gene expression of transforming growth factor-beta1 in bronchoalveolar lavage fluid was examined by using enzyme-linked immunosorbent assay (ELISA). The gene expressions of transforming growth factor-beta1, transcription factor Smad 3 and Smad 7 in lung were analyzed by using immunohistochemical technique (SP) and the image analysis.

RESULTSModel group mainly had Grade III approximately IV silicotic nodule while Qidan group and tetrandrine group had Grade II silicotic nodule. In Qidan group and tetrandrine group, lung/body coefficient and content of hydroxyproline and expression of transforming growth factor-beta1 and Smad3 in lung and expression of transforming growth factor-beta1 in bronchoalveolar lavage fluid were lower than those in model group (P < 0.05). Expression of Smad 7 in lung was higher than model group (P < 0.05). Injury of kidney occurred in tetrandrine group.

CONCLUSIONQidan granules and tetrandrine could inhibit expression of both Smad 7 and transforming growth factor-beta1 and promote expression of Smad 3. Qidan granules and tetrandrine could inhibit remarkably silicotic fibrosis in rats. Qidan granules are safer than tetrandrine.

Animals ; Benzylisoquinolines ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Male ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Silicosis ; drug therapy ; metabolism ; Smad3 Protein ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

The study was purposed to investigate whether the cyclooxygenase inhibitors from some dietary vegetables can inhibit platelet aggregation function by the arachidonic acid (AA). The vegetable juice was mixed with platelet rich plasma (PRP), and asprin was used as positive control. The maximum ratio of platelet aggregation induced by AA was measured on the aggregometer; heme and cyclooxygenase-1 (COX(1)) or cyclooxygenase-2 (COX(2)) were added to test tubes containing COX reaction buffer, the mixture was vortex-mixed and exposed to aspirin or vegetable juice, followed by addition of AA and then hydrochloric acid (1 mol/L) was added to stop the COX reaction, followed by chemical reduction with stannous chloride solution. The concentration of COX inhibitors was detected by the enzyme immunoassay kit; vegetable juice (aspirin as positive control) was mixed with whole blood, which was followed by the addition of AA, and then the reaction was stopped by adding indomethacin, centrifuged, then the supernatant was collected, and the plasma thromboxane B(2) (TXB(2)) was measured by radioimmunoassay. The results showed that spinach juice, garlic bolt juice, blanched garlic leave juice and Chinese leek juice could inhibit by 80% human platelet aggregation induced by AA. 4 kinds of vegetables were all found a certain amount of cyclooxygenase inhibitors, which COX(1) and COX(2) inhibitor concentrations of spinach were higher than that of aspirin; 4 vegetable juice could significantly reduce the human plasma concentrations of TXB(2) induced by AA (p < 0.05). It is concluded that 4 kinds of raw vegetables containing cyclooxygenase inhibitors inhibit the production of TXA(2) and thus hinder platelet aggregation. Raw spinach, garlic bolt, blanched garlic and chinese leek inhibit significantly AA-induced human platelet aggregation in vitro. 4 kinds of vegetables may have a good potential perspective of anti-platelet aggregation therapy or prevention of thrombosis.
Adult ; Arachidonic Acid ; metabolism ; Blood Platelets ; drug effects ; Cyclooxygenase Inhibitors ; pharmacology ; Female ; Humans ; Male ; Platelet Aggregation ; drug effects ; Vegetables ; chemistry