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.

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

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.

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

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

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

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

OBJECTIVETo explore the correlation between ultrasonic scores, routine blood tests and stages of hepatic fibrosis in patients with chronic hepatitis B (CHB), and identify non-invasive indexes to establish a diagnostic model for liver cirrhosis.

METHODSA retrospective analysis of 428 patients with CHB undergoing liver biopsies was conducted. The patients' hematology, serum biochemical indexes, serum alpha fetal proteins (AFP), serum HBeAg status and ultrasonic scores were statistically analyzed. A diagnostic model was established by stepwise discriminant analysis, and aspartate aminotransferase (AST) to platelet ratio index (APRI) was used to estimate the diagnostic value.

RESULTSPartial correlation analysis indicated that platelet, serum albumin, bilirubin, AST, ratio of AST to alanine aminotransferase, prothrombin time and ultrasonic scores were correlated to the stages of liver fibrosis, and significantly differed between patients with and without liver cirrhosis. Logistic regression analysis identified ultrasonic scores, platelet, serum bilirubin, albumin and AST as indexes affecting the diagnosis of compensated cirrhosis. The area under receiver operation curve of model was 0.907. The cirrhosis index (CI) of -0.94 for this model was suitable for screening, with specificity of 85.0%, sensitivity of 81.7%, and accuracy of 84.3%. About 56.2% of the patients' CI was lower than -2.0 with the negative predictive value of 97.0% and the rate of missed diagnosis of 3.0%. About 18.2% of the patients' cirrhosis probabilities were above 0.15, with positive predictive value of 77.3%, and only 2.7% of the patients had mild fibrosis (F2), suggesting that nearly 75% of the patients did not have to receive liver biopsies.

CONCLUSIONThis diagnostic model integrates the ultrasonic scores, platelet, serum bilirubin, albumin and AST to enable effective screening and prediction of compensated cirrhosis, and can reduce the number of patients required to undergo liver biopsy by about 75%.

Adult ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; Female ; Hepatitis B, Chronic ; complications ; Humans ; Liver Cirrhosis ; blood ; diagnosis ; diagnostic imaging ; Logistic Models ; Male ; Platelet Count ; Retrospective Studies ; Sensitivity and Specificity ; Serum Albumin ; analysis ; Ultrasonography

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

OBJECTIVETo test whether nuclear factor kappa B plays an important role in the apoptosis-inhibitory effect of hepatitis B virus (HBV) P22(e) protein.

METHODSHepG2 cells were transfected with recombination plasmid pEGFP-HBVP22(e). The Act-D and TNF alpha were used to induce apoptosis. NF-kappa B inhibitor ALLN were used to inhibit the signaling pathway. The activation of NF-kappa B was EMSA, and the nulear translocation of NF-kappa B was determined by immuno-staining.

RESULTSLaser scanning confocal microscopy and EMSA indicated that HBV P22(e) protein enhanced the nuclear translocation of NF-kappa B after apoptosis induction. ALLN treatment inhibited the nuclear translocation of NF-kappa B, and blocked the apoptosis-inhibiting effect of HBV P22(e) protein.

CONCLUSIONThis study indicates that HBV P22(e) protein inhibits apoptosis of hepatocyte via the NF-kappa B signaling pathway.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B virus ; genetics ; Humans ; Leupeptins ; pharmacology ; Liver Neoplasms ; metabolism ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Plasmids ; Signal Transduction ; drug effects ; Transfection ; Viral Core Proteins ; metabolism