Objective To explore the regulating mechanism of inducible nitric oxide synthase(iNOS) in hepatic injury of obstructive jaundice (OJ) in vivo and in vitro experiments. Methods (1) Rat hepatocytes were isolated by in situ collagenase perfusion and primary culture. Hepatocytes were pretreated with various concentrations of iNOS inhibitor SMT for 20 min. After pretreatment, 50?M GCDC was added for an additional 24hr. Cells were next detected by FCM and TUNEL.(2) Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct in rats. After BDL for 3d、7d、14d、and 21d, the apoptotic status in liver of all rats were determined with TUNEL, and iNOS protein in liver of OJ was ditermined with immunohistochemistry method. Results (1) SMT decreased GCDC-induced apoptosis in a concentration-dependent manner. (2) The apoptotic rate of liver was related to length of time of OJ. Apoptosis index (AI) was highest from rats with 14d bile duct ligation. The stronger the iNOS expression, the higher was the number of apoptotic cells that was found in OJ. Conclusions iNOS is involved in the regulation and the occurrence and progression of hepatic injury of obstructive jaundice.

The effect of targeted magnetic nanoparticles on hepatoma and the underlying mechanism were examined. Nude mice transplanted with a human hepatoma cell line (HepG2 cells) were randomized into 5 groups, including: (1) group A, receiving normal saline, (2) group B, receiving 5-fluorouracil (5-Fu), (3) group C, receiving magnetic nanoparticles containing 5-Fu, (4) group D, consisting of treatment with magnetic nanoparticles containing 5-Fu and inside magnetic field and (5) group E, receiving pure magnetic nanoparticles and inside magnetic field. Morphological features of transplanted tumors in mice in each group were observed under transmission electron microscope (TEM). The expression of bcl-2/bax protein was immunohistochemically detected by SABC method. The results showed that a large number of apoptotic tumor cells were found in group B and group D under TEM. The expression of bcl-2 protein was significantly decreased and the expression of bax protein increased significantly in both group B and D as compared with those in group A, C and E (P<0.01 for all). The decrease in bcl-2 and the increase in bax were more in group D as compared with group B (P<0.01). It is concluded that the targeted magnetic nanoparticles containing 5-Fu can improve the chemotherapeutic effect of 5-Fu by decreasing bcl-2 expression, increasing bax expression and inducing apoptosis of the liver cancer cells.

OBJECTIVETo establish the chromatographic fingerprint for the quality control of gamboge.

METHODAnalysis on a Luna C8 (4.6 mm x 250 mm, 5 microm) column eluted with mobile phases containing acetonitrile and 0.1% glacial acetic acid in water in gradient mode. The flow rate was 1.0 mL x min(-1) and the detection wavelength was at 362 nm. The temperature of column was 25 degrees C. And data of 11 batches of gamboge samples from different sources were analysed by "similarity evaluation for chromatographic Fingerprint of Traditional Chinese Medicine" software.

RESULTThirteen common peaks were selected in chromatograms, and all the common peeks were separated effectively.

CONCLUSIONThe precision, repeatability, and stability of this method were satisfying. The method developed can be used to identify and evaluate the quality of gamboge.

Objective To investigate the experience of surgical treatment of primary extraperitoneal pelvic neoplasms,in order to improve tumor resection rate and safety.Methods The clinical data of 29 cases of primary extraperitoneal pelvic neoplasms were retrospectively analyzed from 1995 to 2013.To evaluate tumor resection preoperatively by CT,MRI and three dimensional reconstruction (3 d) medical technology,5 cases of preoperative interventional vascular embolization,intraoperative 2 cases in the iliac artery ligation,2 cases of intraoperative temporary blocking abdominal aorta.Intraoperative combined a variety of surgical approach to remove the tumor.Results Twenty-nine cases of patients,11 underwent tumor resection,5 underwent tumor resection and rectum resection plus sigmoid colostomy,the structure of 4 underwent tumor resection and repair damaged + sigmoid colon rectum temporary colostomy,3 underwent tumor resection and bladder partial nephrectomy,4 underwent tumor resection and uterine ovarian resection,2 underwent tumor resection + tail sacral tumor resection.25 patients recover well after the surgery,complications of 4 cases:intestinal obstruction in 2 cases respectively,pelvic abscess in 1 case,lower limb venous thrombosis in 1 case,cured by conservative and interventional therapy.No operative mortality and incidence of complications was 13.8％ (4/29).Conclusion Pelvic tumor preoperative imaging evaluation,combined incision and multi-visceral resection,fractional resection and recurrence after resection is the effective examination and treatment.

Objective:To develop a method for the quantitative determination of ambrisentan. Methods: 1 H NMR spectra were obtalned with a Bruker AscendTM 400 superconducting NMR spectrometer. For each sample, DMSO-D6 was used as the solvent, the pulse width was 10. 0 μs, the delay time was 5 s and the scanning time was 16. Results: The proton peaks of ambrisentan at δ6. 16 ppm and maleic acid atδ6. 28 ppm were used as the quantitative peaks. The linear regression equation of peak area and quality ratio was Y=0. 140 7X+0. 034 8 with the correlation coefficient of 0. 999 4. RSD was 0. 2%(n=6)in the repeated experiments. The absolute content of ambrisentan reference substance was 99. 9%. Conclusion: The results showed that 1 H NMR can be used in the quantitative determination of ambrisentan without reference substance. The method is reliable, rapid, accurate and simple.

This study was aimed to establish a HPLC fingerprint of saponins in Sanjie-zhentong Capsule in order to make a quantitative analysis of the quality of Sanjie-zhentong Capsule. The Waters Symmetry ShieldTM RP18 (4.6 mmí 250 mm, 5 μm) column was used with a mobile phase of acetonitrile-water gradient elution. The flow rate was 1.2 mL/min. The column temperature was 30℃. The detection wavelength was 203 nm. The results showed that the fingerprint chromatography included 9 mutual peaks. The similarity among batches was more than 0.95. Compared with reference substance, five characteristic components were recognized. The five components are notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd. It was concluded that this method was rapid, simple and accurate and can be used as one of the effective methods for the quality control of Sanjie-zhen-tong Capsule.

The regulating mechanism in hepatic injury caused by obstructive jaundice (OJ) was examined in this study. Rat hepatocytes were harvestedby in situ collagenase perfusion and subjected to primary culture. The heptocytes were pre-treated with various concentrations of protein kinase C(PKC) agonist PMA and its inhibitor chelerythrine and cultured for 20 min. After the treatment,50 μmol/L glycochenodeoxycholate (GCDC) was added and the cells were cultured for an additional24 h. Cells were then detected by flow cytometry (FCM) and TUNEL. After hepatocytes were treated with different concentrations of fructose and 100 μM GCDC, the cells were examined by FCM and TUNEL. Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct. After BDL, the rats were fed with or without fructos and sacrificed 3, 7, 14 and 21 days after the ligation. The apoptotic status was observed in liver of all rats with TUNEL and PKC protein in liver of OJ was studied by immunohistochemical method. Our results showed that PMA increased GCDC-induced apoptosis and chelerythrine decreased GCDC-induced apoptosis in a concentration-dependent manner. After the treatment with fructose of different concentrations, 100 μM GCDC decreased the apoptotic rate and the apoptotic rate decreased with the increase of fructose concentration. The apoptotic rate of liver was related to the time of OJ. Without the treatment of fructose, PKC and apoptosis index (AI) were highest 14 days after the bile duct ligation. With the treatment of fructose, apoptosis index (AI) and PKC were decreased from the 14th day after the bile duct ligation. It is concluded that PKC is involved in the regulation of apoptosis in the liver cells with OJ and plays important roles in the development and progression of liver injury caused by OJ.Fructose can protect hepatocytes in the bile salt-induced apoptosis by regulating PKC.

Objective To study the effect of magnetic nanoparticles on human cholangiocarcinoma xenograft in nude mice, and compared with otherchemotherapy drugs Methods We established human cholangiocarcinoma xenograft in nude mice with QBC939 cell line.The nude mice were devided into 4 groups randomly.Saline,5-FU, Gemcitabine and magnetic nanoparticles were given to nude mice through tail vein on 20d after implanting QBC939 cell line. Calculations were done at different time after treatment in order to compare tumor volume,inhibition ratio of tumor and tumor growth curve of each group. The nude mice were killed on 35d after treatment to harvest tissue for electron microscopic examination to observe ultra-structural changes. Results The tumor volume of control, 5-FU, magnetic nanoparticles and Gemcitabine groups was (2256.1?267.1) mm3, (2096.5?237.9)mm3,(1392.2?189)mm3, and (1534.9?115 )mm3 respectively.The last two groups have significant difference compared to the first two groups(P

This study was aimed to identify the main related substance in baicalein in order to provide basic study data for the safety, efficacy and quality control of the medicine. The analysis was carried out on activated carbon and silica gel column. The compound structure was identified by spectral analysis, which included UV, IR, NMR and MS. The results showed that the related substance was identified as oroxylin A. It was concluded that the determina-tion provided scientific data for studying the impurities of baicalein and improving its quality.

The effect of targeted magnetic nanoparticles on hepatoma and the underlying mechanism were examined. Nude mice transplanted with a human hepatoma cell line (HepG2 cells) were randomized into 5 groups, including: (1) group A, receiving normal saline, (2) group B, receiving5-fluorouracil (5-Fu), (3) group C, receiving magnetic nanoparticles containing 5-Fu, (4) group D,consisting of treatment with magnetic nanoparticles containing 5-Fu and inside magnetic field and (5)group E, receiving pure magnetic nanoparticles and inside magnetic field. Morphological features of transplanted tumors in mice in each group were observed under transmission electron microscope(TEM). The expression of bcl-2/bax protein was immunohistochemically detected by SABC method.The results showed that a large number of apoptotic tumor cells were found in group B and group D under TEM. The expression of bcl-2 protein was significantly decreased and the expression of bax protein increased significantly in both group B and D as compared with those in group A, C and E(P＜0.01 for all). The decrease in bcl-2 and the increase in bax were more in group D as compared with group B (P＜0.01). It is concluded that the targeted magnetic nanoparticles containing 5-Fu can improve the chemotherapeutic effect of 5-Fu by decreasing bcl-2 expression, increasing bax expression and inducing apoptosis of the liver cancer cells.