In order to evaluate the value of magnetic resonance cholangiography (MRC) and dynamic examination of duodenal fluid in the differential diagnosis between extrahepatic biliary atresia (EHBA) and infantile hepatitis syndrome (IHS), 52 patients with infantile cholestatic jaundice were examined by MRC and duodenal fluid examination. Original interpretations were compared with clinical outcome. Calculated sensitivity of duodenal fluid examination in diagnosis of EHBA was 100%, and specificity was 91.1%. Sensitivity of MRC in the diagnosis of EHBA was 94.4% and specificity 88.24%. The sensitivity of MRC and examination of duodenal fluid combined in diagnosis of EHBA was 94.4% and specificity 97.06%. We are led to conclude that MRC and dynamic examination of duodenal fluid are useful in the differential diagnosis between IHS and EHBA and the combined use of the two techniques yield better results.

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

The content of carboxylate ions in marine sulfate polysaccharide 911 was determined by conductometric titration. In the mean while, the sample was transformed with cation exchange resin,the molar ratio of sufate ions to carboxylate ions was determined. The results showed that conductometric titration is a simple,rapid and accurate method.

Objective To develop a method for simultaneous determination of three hydrophilic components and two lipophilic components in Radix et Rhizoma Salviae Miltiorrhizae. Methods The RP-HPLC method was performed by using a Welchrom C18 column(250 mm×4.6 mm,5 μm)with a mobile phase of acetonitrile (A)-0.1%phosphoric acid(B). The gradient elution program was as follows:0-15 min,10%→12%A;-35 min,12%→20%A;-45 min,20%→60%A;-65 min,60%→65%A;-80 min,65%→80%A;-90 min,10%A. The flow rate was kept at 1.0 mL?min-1 . The detection wavelength was set at 280 nm. The column temperature was 30 ℃ . Results A good linearity was obtained over 0.059 5-2.380 0 μg for tanshinol, 0.346 0-13. 840 0 μg for rosmarinic acid, 0. 656 0 - 26. 240 0 μg for salviamolic acid B, 0. 420 0 - 16.800 0 μg for cryptotanshinone and 0.414 0- 16.560 0 μg for tanshinoneIIA, respectively ( r = 0.999 9). The average recovery rates were between 98.69%-100.91% with RSD less than 1.2%(n = 6). Conclusion The method is rapid, accurate, credible and repeatable, and can provide basis for the quality control of Radix et Rhizoma Salviae Miltiorrhiza.

Objective To evaluate endoscopic sphincterotomy(EST)for the management of choledocholithiasis. Methods Between July 1987 and March 2007,991 cases of choledocholithiasis treated with EST were reviewed,and 710 cases were followed-up. Results All cases were treated with EST.The common bile duct stones were removed in 909 cases(91.7%,909/991).The rate of complications after EST was 6.0%(59/991)including pancreatitls in 29 cases(2.9%),hemorrhage in 18 cases(1.8%),cholangitis in 11 cases(1.1%),and duodenal perforation in 1 case(0.1%).There was no mortality in this group.Follow-up found reflux cholangitis in 51 cases(7.2%)and recurrent choledocholithiasis in 42 cases(5.9%). Conclusion Therapeutic endoscopy for choledocholithiasis is safe and effective.

Background:Recently,cancer stem cells( CSCs)has become a hot topic in cancer research. For this research topic,the primary problem is the isolation and identification of CSCs,and suspension culture is considered to be an important method for CSCs isolation. Aims:To study the isolation and identification of gastric cancer stem cells( GCSCs) and evaluate its stemness characteristics. Methods:GCSCs-like cells were enriched through spheroid formation by suspending the human gastric cancer cell lines MKN45,MGC803 and SGC7901 in serum-free medium containing epidermal growth factor( EGF)and basic fibroblast growth factor( bFGF). The third generation tumor spheres obtained were tested for the ability of colony forming;its stemness markers were identified by immunofluorescent staining,and migration and invasion capabilities were assessed by wound scratch test and Transwell assay,respectively. Growth inhibition of tumor spheres in response to cisplatin was assessed by CCK-8 assay. Results:High expressions of stem cell markers CD44 and CD54 were observed in the tumor spheres,but gastric parietal cell marker H+ /K+-ATPase was lowly expressed. Compared with parental gastric cancer cells,tumor spheres had stronger colony forming ability(P<0. 05);Furthermore,healing of wound scratch was accelerated(P<0. 05),and number of invasive cells in Transwell chamber was increased in tumor spheres(P<0. 05). The 50% inhibition concentration of cisplatin to tumor spheres increased significantly when compared with parental gastric cancer cells. Conclusions:GCSCs enriched tumor spheres can be obtained by suspension culture with serum-free medium. The tumor spheres possess enhanced capabilities of self-renewal,proliferation,migration,invasion and potential of multilineage differentiation;meanwhile,it is more resistant to chemotherapy.

Objective To study PTEN protein expression and clinical significance in patients with diffuse large B cell lymphoma. Methods Immunohistochemical staining was used to determine the PTEN protein expression in 40 cases of primary diffuse large B lymphoma tissuse. The results were analyzed by Kaplan-Merie survival analysis, Log-Rank test and Logistic regression analysis. Results PTEN protein was positive in 16 cases and negative in 24 cases. There was no significant difference between two groups in twoyear overall survival rate(62.5 % vs 66.7 %, P >0.05). Survival analysis showed that patient' s survival time gradually were reduced with extended time between PTEN protein-positive group and negative group, lower in PTEN-positive group than the negative group, but there was no significant difference in survival curve (P >0.05) in the two groups. We compared characteristics of patients between PTEN protein positive and negative groups,including molecular type, patient' s age, stage, LDH, physical score and extranodular invasion, there was no significant difference among them. PTEN protein was not correlated with prognosis, while International Prognosis Index(IPI) was still a risk factor (OR >1). Conclusion PTEN protein expression may not predict the outcome in diffuse large B cell lymphoma, but IPI still is a predictor.

OBJECTIVE:To establish a method for content determination of schizantherin E,gomisin J,angeloylgomisin H,schisantherin A,schisantherin B,schisanhenol,anwuligan,schizandrin A,schizandrin B and schizandrin C in Wuzhi tablets.METHODS:RP-HPLC method was adopted.The determination was performed on Symmetry C18 column with acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set 225 rm,and the column temperature was 30 ℃.The sample size was 10 μL.RESULTS:The linear ranges of schizantherin E,gomisin J,angeloylgomisin H,schisantherin A,schisantherin B,schisanhenol,anwuligan,schizandrin A,schizandrin B and schizandrin C were 2.25-67.5ng(r=0.999 6),2.1-63 ng(r=0.999 8),28-840 ng(r=0.999 9),124.6-3 738 ng(r=0.999 9),22.7-681 ng(r=0.999 9),32.7-981 ng(r=0.999 9),47-1 410 ng(r=0.999 9),208-6 240 ng(r=0.999 9),5.36-160.8 rig(r=0.999 9),4.48-134.4 ng(r=0.999 8).The limits of quantitation were 14.17,13.32,9.33,11.37,14.62,19.88,14.66,12.50,16.40,13.55 rg.The limits of detection were 4.62,4.60,3.08,3.76,4.81,6.74,4.93,4.16,5.86,5.03 ng.RSDs of precision,stability and reproducibility tests were less than 3.0％;the recoveries were 96.36％-100.00％(RSD=1.83％,n=6),95.00％-100.00％(RSD=2.07％,n=6),95.00％-98.00％(RSD=1.22％,n=6),95.37％-98.91％ (RSD=1.29％,n=6),95.62 ％-103.71％ (RSD=2.85％,n=6),97.33％-102.67％ (RSD=2.00％,n=6),95.00％-99.33％ (RSD=1.75％,n=6),97.24％-104.93％ (RSD=2.63％,n=6),95.00％-97.50％ (RSD=1.42％,n=6),96.00％-102.00％ (RSD=2.45％,n=6),respectively.CONCLUSIONS:The developed method is accurate,sensitive and reproducible,and it can be used for content determination of 10 lignanoids in Wuzhi tablets.

Objective To discuss the clinical and pathological characteristics, treatment results and prognosis of primary parotid malignant lymphoma. Methods Pathological subtypes, clinical stages and treatment of the 24 patients with primary parotid malignant lymphoma were retrospectively analysed. Kaplan-Meier method was used in the survival analysis and Log-Rank method was used in the statistic study. Results The 5-year progression free survival (PFS) and overall survival (OS) were 79.2 % and 83.3 %, respectively. The 5-year PFS and OS were 89.5 % and 94.7 % for 19 patients with low-grade malignant lymphoma (including MALTL and Ⅰ/Ⅱ grade FL). The differences of the 5-year PFS and OS of 9 patients received chemotherapy and of 10 patients with on chemotherapy had no statistical significance. Conclusion The incidence of primary parotid malignant lymphoma is low, at earlier clinical stage, and most of its pathological subtype were B-cell low-grade malignant non-Hodgkin lymphoma. Surgery and/or radiotherapy should be the first choice for patients with early stage low-grade malignant lymphoma, whereas combined modality therapy was probably the best choice for patients with DLBCL.

Objective To determine the composition of abnormal red cell membrane skeletin. Methods By sodium dodecyl sulphate polyacrylamide gel electrophoresis of ghostcorpuscles, we quantified the amount of protein by densitometric evaluation.Results The results showed that in β-thalassernia, the amount of spectrin, 4.5 protein and globin significantly increased compared with the controls (26.05 + 1.46, 21.69 + 1.86; 22.87 + 5.61, 12.99 +2.33; 15.23+3.31 and 4.97+2.73, respectively, P＜0.05).Conclusion These data suggest that the erythrocyte membrane protein composition matched with globin in patients with β-thalassemia of the racial Li minority are different from the normal control. These factors increase rigidity but decrease deformability of the β-thalassemic red cell membrane, which may lead to hemolytic anemia.