Objective In order to discuss the surgical treatment of iatrogenic bile duct injury.Methods 156 cases of iatrogenic bile duct injury during recent 20 years were reviewed and analyzed in this paper.Results The total re-operation rate was 85.71% (126/147), the re-operation rate of LC, OC, OC plus exploration of common bile duct were 95.24%, 87.27%, 62.50% respectively (P

Objective To study the effects of various media, salt intensity, and organic components on the growth of Salvia miltiorrhiza adventitious roots and the synthesis of tanshinone ⅡA and protoca-techuic aldehyde. MethodsThe adventitious roots were obtained through tissue culture by manipulation of various media, salt intensity, and organic components and the contents of tanshinone ⅡA and protocatechuic aldehyde were determined by HPLC. ResultsThe effect of media MS, LS, B5, White, and SH on adventitious roots of S. miltiorrhiza was observed. Adventitious roots grew better under high salt intensity while secondary metabolite biosynthesis was accelerated under low salt intensity in MS basal medium.The reciprocity of five organic components had significant effect on root growth; glycin favored the synthesis of tanshinone ⅡA; scarcity of one of inosital, glycin, VB1, and VB6 inhibited the synthesis of protocatechuic aldehyde. ConclusionMS Basal medium is used for adventitious root culture. The results show that salt intensity and organic components have significant effects on adventitious root culture of S. miltiorrhiza and secondary melabolite synthesis.

Objective To study the effects of carbon, nitrogen, and phosphate sources on the growth of Salvia miltiorrhiza adventitious roots and the contents of tanshinone ⅡA and protocatechuic aldehyde. Methods The adventitious roots were obtained through tissue culture by manipulation of carbon, nitrogen, and phosphate sources and the contents of tanshinone ⅡA and protocatechuic aldehyde were determined by HPLC. Results Carbon, nitrogen, and phosphate sources were necessary for the culture of S. miltiorrhiza adventitious roots. The highest times of root multiplication were achieved at sucrose level of 30 g/L after 20 d culture, 60 g/L sucrose and low level sucrose were favorable for biosyntheses of tanshinone ⅡA and protocatechuic aldehyde, respectively. The highest root yield and tanshinone ⅡA content on day 25 were obtained by intermittent sugar adding during cultivation, and the production of adventitious roots and tanshinone ⅡA were 2.3-and 2.4-fold compared with those of control, respectively. The maximum root growth rate, contents of tanshinone ⅡA and protocatechuic aldehyde were achieved while NH4+-NO3-was 1∶4, 1∶4, and 1∶1, respectively when concentration of total nitrogen source was kept at 60 mmol/L. To compare with the control group, changing of KH2PO4 concentration could favor for the adventilious root growth, but high KH2PO4 concentration inhibited tanshinone ⅡA biosynthesis. ConclusionThe results show that various carbon, nitrogen, and phosphate sources have the significant effects on adventitious root culture of S. miltiorrhiza. The best carbon source and its concentration, nitrogen and phosphate sources for the growth of S. miltiorrhiza adventitious root and the synthesis of secondary metabolite are confirmed.

Objective To construct a mammalian vector encoding angiostatin kringle 5 (K5) under the control of ?-fetoprotein (AFP) enhancer and albumin promoter, and to observe the expression of angiostatin by introducting angiostatin gene into hepatocellular carcinoma cells through gene transfection. Methods Angiostatin cDNA was amplified from normal human eukaryotic cells by using RT-PCR. Meanwhile, AFP enhancer and albumin promoter sequences were directed cloned and were inserted into vector pcDNA3.1. The recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was constructed, which contained the angiostatin K5 cDNA sequence that was under the control of the AFP enhancer and promoter. Angiostatin K5 cDNA was introduced into human AFP positive hepatocellular carcinoma cell lines with the transfected cultured cells that were mediated with Lipofectamine 2000. The expression of angiostatin K5 was analyzed by Western blot and the protein was dectected with anti-His antibody. Results The 500-base pair of angiostatin K5 was in accordance with the expected sequence and the recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was also confirmed as the anticipated sequence. The expression of angiostatin K5 in AFP positive hepatocellular carcinoma cells was detected both by SDS-PAGE and Western blot. Conclusion Efficient construction and expression of angiostatin K5 to AFP positive cells make it possible for antiangiogenesis therapy of human hepatocellular carcinomas, which may provide a promising approach.

Objective To determine quercetin content in Polygonum chinense L.from different growing areas.Methods RP-HPLC was used to determine quercetin content.Chromatographic conditions were as follows:chromatographic column of Agilent TC-C18 (4.6 mm?250 mm,5 ?m),mobile phase consisting of methanol-0.6 %phosphoric acid (52:48),with a velocity of 1 mL?min-1,column temperature at (25?1)℃,detection wavelength being 365 nm,elution time of 20min,and an injection volume of 20 ?L.Results Standard curve regression equation of quercetin is:Y=85 355X-48.16(r=0.999 3).A good linearity was showed in the range of 0.004～0.012 mg?mL-1,with average recovery being 100.3 %,and RSD of 1.44 %.The content of quercetin in the samples from different growing areas is in the range of 183.0～848.1 ?g?g-1.Conclusions The method is simple and reliable,and can be used to control the quality of Polygonum chinense L.The existence of some differences of quercetin content between medicinal materials from different growing areas indicates that we should make a quality standard to ensure the safety and effectiveness of Polygonum chinense L.in clinic.

Objective To study the therapeutic effect and mechanism of Dicliptera chinensis polysaccharide ( DCP) on liver injury induced by antituberculosis drugs. Methods Sixty mice were randomly divided into six groups, namely normal control group, model group, glucurolactone group (in the dosage of 200 mg·kg-1·d-1), and high-, middle- and low-dose DCP groups ( in the dosage of 600, 400, 200 mg·kg-1·d-1, respectively). Except for the normal control group, the rats in the other groups were given intragastric administration of isoniazid and rifampicin ( 100 mg/kg) to induce liver injury model, and were simultaneously treated with corresponding agents, once a day. On the experiment day 30, the blood and liver tissue were sampled. The serum levels of alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST) , alkaline phosphatase ( AKP) and microsomal nitric oxide ( NO) were detected by biochemical method. The contents of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) in liver tissue were determined by enzyme-linked immunosorbent assay ( ELISA) , and the hepatic histopathology was observed after HE staining. Results In DCP groups, the hepatic pathological changes of the mice were improved, the number of the inflammatory cells was reduced, and the activities of serum ALT, AST and AKP as well as the contents of hepatic TNF-α, IL-6 and NO were reduced ( P<0.05 or P<0.01 compared with those in the model group). Conclusion Dicliptera chinensis polysaccharide is effective for liver injury induced by antituberculosis drug, and the mechanism may be associated with its anti-inflammatory action.

Aim Based on the effect of total flavonoids extracted from Polygonum perfoliatum L.(TFP) against dimethylnitrosamine(DMN)-induced hepatic fibrosis(HF), to investigate the anti-fibrotic mechanism of TFP.Methods Ninety SD rats were divided into normal group, model group, colchicines(0.1 mg·kg-1) group, and TFP(200, 100, 50 mg·kg-1) group.Except the rats of normal group, other rats were injected intraperitoneally with volume fraction 0.5% DMN solution(2 mL·kg-1) for eight weeks, once every two days.From the first day of modeling, each administration group was given the corresponding dose of drugs to intervene, and the normal group and model group were given an equal volume of solvent, once a day.At the end of the eighth week, the blood and liver tissues were collected.Liver tissue was taken at a fixed position, and the degree of liver tissue was observed by HE staining.The contents of serum ALT, AST, SOD and MDA were measured using colorimetric method;the levels of serum HA, LN, PCⅢand Ⅳ-C were detected using enzyme-linked immunosorbent assay(ELISA);the levels of TNF-α, IL-1β and IL-6 in liver tissues were detected by ELISA;the expression of α-SMA, TGF-β1, p-JAK2 and p-STAT3 were detected by Western blot.Results Compared with the model group, TFP(200, 100, 50 mg·kg-1) could improve the liver tissue lesions, reduce the expression of ALT, AST, HA, LN, PCⅢ, IV-C and MDA, increase SOD activity, reduce the levels of TNF-α, IL-1β and IL-6, and inhibit the expression of α-SMA, TGF-β1, JAK2, STAT3, p-JAK2 and p-STAT3.Conclusion TFP could inhibit DMN-induced HF of rats, which may be involved with antioxidant and inhibiting expression of TGF-β1, JAK2/STAT3 signaling pathway and inflammatory response.

With the continuous development of the cancer stem cell theory,biomarkers for CSCs (Cancer stem cells) have gradually become the hot issues in tumor research.As the most commonly used CSCs biomarker,CD44 gains lots of attentions.This paper will review recent research about CD44 molecule and its powerful role in regulating tumor progenesis and progression.

Aim To investigate the effect of DHA on NGF-induced neuronal differentiation of PC12 cells and explore the possible mechanism via regulating BMP pathway. Methods PC12 cells were treated with 100μg·L-1 NGF and 100 μg·L-1 NGF + 10 μmol· L-1 DHA for 3, 6 and 9 days respectively. The length and number of neurite were detected by immunofluores-cenc. DHA content was analyzed by gas chromatogra-phy in all groups. The protein expression of BMP4, BMP7 , BMPR-II and p-Smad 1/5/8 was determined by Western blot. Results The length of total primary neurite in NGF+DHA groups was obviously increased, longer than that in NGF group; DHA content in 10μmol · L-1 DHA group was higher than that in the control group;NGF+DHA groups also unregulated the protein expression of BMP4 , BMP7 , BMPR-II and p-Smad 1/5/8 . Conclusion DHA promotes NGF-in-duced neuronal differentiation in PC12 cells, which may be associated with the upregulation of BMP path-way protein.