A novel method for the simultaneous quantification of seven metabolites of polycyclic aromatic hydrocarbon in human urine was developed using online solid phase extraction-HPLC with double ternary liquid chromatography system combined with fluorescence detector. The target compounds were online concentrated on the Turboflow Cyclone solid phase extraction column at first, then transferred by the six-way valve to the Hypersil Green PAH column for separation with acetonitrile and water as mobile phase at a flow rate of 1. 0 mL/min and at 35 ℃. A single sample analysis cycle took only 20 min. Under the optimized chromatographic conditions, the method showed good linear relationship ( r≥0. 999 ) in the range of 5-2000 ng/L or 50-20000 ng/L. The LODs were 0. 5-15 ng/L, and the recoveries were 80. 7%-110. 7%. The proposed method was successfully applied in the detection of metabolites of polycyclic aromatic hydrocarbons in urine from several smokers and non-smokers. The concentrations of 2-hydroxynaphthalene, 1-hydroxynaphthalene, 2-hydroxyfluorene, 2-hydroxyphenanthrene, 4-hydroxyphenanthrene and 6-hydroxychrysene in the smokers urine were much higher than that in non-smokers.

Background Epidemiological investigation in human has been conclusive. In postmenopausal women,the incidence of cataract is higher than men at the same age. In addition,hormone replacement therapy may protect against the development of cataract. However,this role of androgen is not clear. Objective This study was to explore the effects of estrogen and androgen on anti-oxidative ability of lens after ovariectomy. Methods Fifty-six three-month-old clean female Wistar rats were randomly divided into normal control group,sham operation group, castration group,estrogen eyedrops group;estrogen injection group;androgen eyedrops group;androgen injection group and 8 rats for each. Ovariectomy was performed in the rats of castration group and gonadal hormone application group, and estradiol benzoate solution or testosterone propionate solution were utilized topically or systemly in 5 months after ovariectomy for 6 weeks respectively. Only abdominal cut was curried out in sham operation group. The lenses of rats were examined weekly under the slit lamp. The serum estrogen and androgen levels of rats were detected before,after operation and 6 weeks following the administration of gonadal hormone. The contents of superoxide dismutase( SOD) , glutathione( GSH) ,malondialdehyde( MDA) and water-soluble protein ( WSP) in rat lens homogenate were detected at the end of the experiment. Utilization of animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results No opacity of lenses was found during the experiment duration in various groups. The serum estradiol levels of rats in sham group were insignificantly different from normal groups in various time points( P>0. 05). The evident decline of serum estradiol was detected in the rats of castration group and gonadal hormone application groups compared with sham group in 5 months after operation( all P<0. 01). However,at the sixth weeks after the system use of estradiol or testosterone,the serum estradiol levels were significantly higher than the castration group and topical application groups of gonadal hormone(P<0. 01). The contents of SOD,GSH and WSP in lenses were considerably increased,but the MDA level in lenses was decreased after system use of estrogen ( P<0. 01). The activity of SOD and GSH were lower after system use of testosterone in comparison with castration rats ( P < 0. 05 ). Conclusion Estrogen can protect lens against oxidation damage. However, androgen, to a certain extent, may contribute to the development of oxidative damage in OVX female rats.

Objective To observe the expression of estrogen receptor(ER)? and ER? in the vaginal wall of women with anterior vaginal prolapse,and to investigate the relationships of ER subtypes with the development of pelvic organ prolapse(POP).Methods Seven premenopausal women and 33 postmenopausal women with anterior vaginal prolapse who underwent surgery in our hospital from July 1999 to July 2004 were analyzed.Nine premenopausal and 8 postmenopausal women with squamous carcinoma of cervix who underwent surgery served as controls.The expression of ER? and ER? in squamous epithelium (SE),lamina propria(LP)and muscular layer(ML)of anterior vaginal wall were studied by immunohistochemical staining.Results(1)Both ER? and ER? were expressed in SE,LP,ML of vaginal wall of premenopausal and postmenopausal women.(2)The expression of ER? was not significantly different in premenopausal and postmenopausal women;the expression of ER? was not significantly different in premenopausal and postmenopausal women with POP;however,it was decreased in postmenopausal women without POP(P

OBJECTIVETo summarize the clinical experience and the role of hepatectomy with portal vein resection and reconstruction hilar cholangiocarcinoma.

METHODSFrom 1998 to 2003, the clinical records of 118 cases with hilar cholangiocarcinoma were reviewed.

RESULTSOf the 118 patients, 66 were performed palliative treatment; and 52 patients underwent radical resection, of which 47 patients, including 11 cases combined with portal vein resection and reconstruction, underwent hepatectomy. The rate of postoperation complication was 22.9% and 27.3% in hepatectomy with or without portal vein resection and reconstruction respectively. The 1, 3-year survival rate were 85.7%, 31.4% and 81.8%, 27.8% in hepatectomy with or without portal vein resection and reconstruction respectively (P > 0.05). Only 5 patients were alive more than 3 years (7.58%), and no patient with palliative treatment lived over 5 years.

CONCLUSIONSPortal vain invasion is not the contraindication of resection for hilar cholangiocarcinoma. Hepatectomy with portal vein resection and reconstruction may raise the radical resection rate of hilar cholangiocarcinoma and improve the results of prognosis.

Adult ; Aged ; Bile Duct Neoplasms ; pathology ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; pathology ; surgery ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Portal Vein ; pathology ; surgery ; Prognosis ; Reconstructive Surgical Procedures ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo study the effect of Feiji Recipe (FR) intervening indoleamine 2,3-dioxygenase (IDO) induced immune escape on the murine model of Lewis lung carcinoma. Methods Totally 48 C57BL/6 mice inoculated with Lewis lung cancer cells transfected with human (enhanced green fluorescent protein,EGFP)-IDO gene were divided into four groups according to radom digit table, i.e., the model group (administered with normal saline by gastrogavage) , the Chinese medicine group (treated with FR Decoction at the daily dose of 100 mg/g by gastrogavage), the 1-methyl-D-trytaphan (1-MT) group (administered with 1-MT mixed liquor at the daily dose of 100 mg/kg by gastrogavage), and the Paclitaxel group (treated with Paclitaxel at the daily dose of 15 mg/kg by peritoneal injection), 12 in each group. The intervention was started from the 2nd day of modeling. The survival time was observed in 24 of them. Ratios of CD4+ CD25+ FoxP3+ regulatory T cells (Treg) in the spleen were detected in the rest 24 mice by flow cytometry respectively.

RESULTSCompared with the model group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells remarkably decreased in the Chinese medicine group, the 1-MT group, and the Paclitaxel group (P < 0. 01). Compared with the Paclitaxel group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells decreased significantly in the 1-MT group (P < 0.05).

CONCLUSIONFR could inhibit the proliferation of lung cancer cells and immune eseape, improve the immune function, and prolong the survival of tumor-bearing mice.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Carcinoma, Lewis Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; Lung Neoplasms ; Mice ; Mice, Inbred C57BL ; Paclitaxel ; T-Lymphocytes, Regulatory

Animals ; Escin ; pharmacology ; Intestines ; blood supply ; Lipid Peroxidation ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; prevention & control

OBJECTIVETo develop a RP-HPLC method for determination of three alkaloids in total alkaloids exfracted from Lotus plumule.

METHODA Hypersil C18 column was used with the mobile phase of methanol-phosphate (73:27, pH 9.0) at the detection wavelength of 230 and 286 nm. The flow rate was 1 mL min(-1).

RESULTThe linear range was 20-700 microg mL(-1), the recovery of three alkaloids were 97.9%, 98.4% and 98.1%. The RSD of intra-day and inter-day was 0. 24% -4. 83%. Different groups of samples were determined by this method. The result showed that the extraction method with methanol was prior to the extraction method with alcohol.

CONCLUSIONThe method is simple, rapid and suitable for the determination of three alkaloids in total alkaloids exfracted from L. plumule.

Alkaloids ; chemistry ; isolation & purification ; Benzylisoquinolines ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Isoquinolines ; analysis ; Loteae ; chemistry ; Phenols ; analysis ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Seeds ; chemistry

To observe the morphological characteristics and hematopoietic function of bone marrow megakaryocyte (MK) in children patients with idiopathic thrombocytopenic purpura (ITP), and to preliminary analyse the cause and mechanism of thrombocytopenia. CD41 McAb immunohistochemical technique was used to detect micromegakaryocyte in bone marrow smear. Plasma clot culture and CD41 McAb immunohistochemical technique were used for the MK-colony forming assay. The results showed that there was no statistical difference of the positive rate of micromegakaryocyte between groups of ITP and control, but type I lymphocyte-like micromegakaryocyte was infrequent. The number of micromegakaryocyte and the formation rates of CFU-MK and BFU-MK in ITP group were significantly higher than those in control group. The normal MK releasing platelet could be easily found in the culture system. The MK colony formation rate was decreased in a patient with chronic ITP. In conclusion, the increment of type II, III, IV micromegakaryocytes is one of pathologic phenomenon of ITP. These small megakaryocytes can develop and mature to normal megakaryocytes in the condition of ex vivo culture. The developmental abnormity of MK is a possible reason for thrombocytopenia among partial patients with ITP, especially the chronic cases.
Adolescent ; Bone Marrow Cells ; pathology ; physiology ; Child ; Child, Preschool ; Female ; Hematopoiesis ; Humans ; Immunohistochemistry ; Infant ; Male ; Megakaryocytes ; pathology ; physiology ; Platelet Membrane Glycoprotein IIb ; analysis ; Purpura, Thrombocytopenic, Idiopathic ; pathology ; physiopathology

OBJECTIVETo investigate the correlation between the stage of hepatic fibrosis and ultrasonographic findings of the liver, spleen and gallbladder and establish a sensitive ultrasonographic semi-quantitative scoring system for screening compensated liver cirrhosis.

METHODSTotalling 248 patients with chronic hepatitis B and hepatitis C virus infection underwent liver biopsy and ultrasonic examination. The images of the liver surface, parenchymal echo, intrahepatic vessels, gallbladder, spleen and diameter of portal vein were analyzed.

RESULTSThe stages of hepatic fibrosis were not correlated to ultrasonographic findings of the liver surface or diameter of portal vein, but hepatic fibrosis of different stages showed significant differences in parenchymal echo, intrahepatic vessels, gallbladder and splenomegaly. In cases with normal liver parenchymal, intrahepatic vessels, gallbladder and spleen, the negative predictive value of the ultrasonographic semi-quantitative scoring system for diagnosing compensated liver cirrhosis amounted to 96.3%. The sensitivity of a score not lower than 5 was 90% for detecting compensated cirrhosis. With a score not lower than 7, the diagnostic accuracy and specificity was 85.9% and 95.2%, respectively, but the sensitivity was lowered to 37.5%.

CONCLUSIONThe ultrasonic images of the liver parenchyma, intrahepatic vessels, gallbladder and spleen in patients with compensated liver cirrhosis vary significantly in patients with hepatic fibrosis of different stages, and this ultrasonographic scoring system allows for a sensitive diagnosis of compensated cirrhosis.

Female ; Fibrosis ; Gallbladder ; diagnostic imaging ; Hepatitis B, Chronic ; complications ; Hepatitis C ; complications ; Humans ; Liver ; diagnostic imaging ; pathology ; virology ; Liver Cirrhosis ; complications ; diagnosis ; Male ; Reproducibility of Results ; Sensitivity and Specificity ; Spleen ; diagnostic imaging ; Splenomegaly ; diagnostic imaging ; Ultrasonography ; methods

OBJECTIVETo explore the effect of ITF2357, a novel histone deacetylase (HDAC) inhibitor, on the growth, differentiation and apoptosis of acute myeloid leukemic (AML) cells and its mechanism.

METHODSAML cell lines kasumi-1 cells as a model for AML1-ETO positive, and THP1 cells for AML1-ETO negative, the leukemic cells proliferation was analyzed by MTT assay, expression of myeloid-specific differentiation antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining and flow cytometry. AML1-ETO, acetyl-histone, and caspase protein was analyzed by Western blot.

RESULTS0.5 µmol/L ITF2357 treatment significantly inhibited kasumi-1 cells proliferation, with the 48 h half inhibitory concentration (IC(50)) of 0.1 µmol/L. The initial inhibitory concentration of THP1 cell line was 5 µmol/L. ITF 2357 induced apoptosis of kasumi-1 cells in a time- and dose-dependent manner. A dose-dependent increase in early apoptosis occurred at 24 hours treatment and in late apoptosis at 48 hours treatment by ITF2357. Early apoptosis cells increased from (1.44 ± 1.52)% to (24.51 ± 5.79)%. Late apoptosis cells increased from (2.37 ± 2.8)% to (63.66 ± 1.56)%. ITF2357 induced AML1-ETO degradation by caspase-dependent pathway. 0.25 µmol/L ITF2357 induced a time- and dose-dependent increase in expression of myeloid cell surface protein CD13 and CD15. 5 µmol/L ITF2357 blocked the cells at G(0)/G(1) phase, G(0)/G(1) cells increased from (39.69 ± 6.56)% to (79.2 ± 6.51)% and s-phase cells declined from (60.12 ± 3.29)% to (18.97 ± 6.62)%. Kasumi-1 cells incubated with 0.5 µmol/L of ITF2357, AML1-ETO protein began to decrease at 24 hours and could hardly be detected at 96 hours. ITF2357 induced AML1/ETO degradation through a caspase-dependent mechanism. At the same time, acetylated H3 and H4 increased.

CONCLUSIONLow-dose HDAC inhibitor ITF2357 can effectively inhibit the AML cells proliferation, especially for AML1-ETO positive AML cells. It inhibits Kasumi-1 cells proliferation degradation of AML1-ETO protein expression, blocks the cells at G(0)/G(1) phase, and induces apoptosis and differentiation of the cells.

Acetylation ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Histone Deacetylase Inhibitors ; pharmacology ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; Oncogene Proteins, Fusion ; metabolism