This paper focuses mainly on the study of optimal fermentation conditions of S-adenosyl-L-methionine.Effects of carbon sources,nitrogen sources,inorganic constituents,growth factors and adding time of L-methionine on the yield,the content and biomass of S-adenosyl-L-methionine are studied.And ingredients of the culture medium are also optimized by the method of uniform design.The final optimum culture medium contains: glucose 30 g,Yeast powder 11 g,(NH_4)_2SO_4 12 g,K_2HPO_4?3H_2O 5 g,KH_2PO_4 10 g,MnSO_4?H_2O 0.09 g,ZnSO_4?7H_2O 0.14 g,MgCl_20.5 g,CaCl_2 0.3 g,CuSO_4 0.005 g per liter. Using that optimum culture medium,the yield of S-adenosyl-L-methionine can reach 0.9 g/L in Erlenmeyer flask which is 30 % higher than before.Experiment on 5 L fermenter reveals that the accumulation of S-adenosyl-L-methionine can reach 2.66 g/L.Biomass is 23.4 g/L.

Objective To compare the reproducibility and reliability of facial soft tissue landmarks using a direct and an indirect placement methods in facial three-dimensional (3D) soft tissue assessment.Methods 3D coordinates of 37 soft tissue landmarks were obtained respectively in 35 normal healthy volunteers from 3D optical capture system and Materialise's interactive medical image control system (Mimics).Landmarks were affixed on the cutaneous surface (direct method) and marked on the 3D reconstructed craniofacial model (indirect method).Intra-class correlation coefficients (ICCs) and paired t-tests were used to compare the reliability and reproducibility of facial landmarks between two placement methods.Results For the direct placement method,86.5 ％ landmarks had an ICC higher than 0.75.For the indirect placement method,75.7％ landmarks had an ICC higher than 0.75.The inter-observer variability using the direct placement method was (1.259±0.566) mm,which was significantly smaller (P＜0.05) than (1.420±0.638) mm obtained by the indirect placement method.Conclusions Compared with indirect method,direct method has higher reliability and reproducibility in evaluating facial soft tissue.

Objective To observe the efficacy of Sorafenib in preventing and treating tumor recurrence after liver transplantation for patients with primary hepatic carcinoma beyond Milan criteria.Methods From March 2008 to June 2010,30 patients of liver transplantation with primary hepatic carcinoma exceeding Milan criteria were randomized into 2 groups,each group of 15 cases.Beginning one month posttransplantatoin patients in the experimental group received oral administration of Sorafenib (400 mg bid),while those in the control group received Capecitabine ( 1500 mg bid) for 14 days every 4 weeks.Drug was withdrawn in patients without recurrence in 18 months after transplantation,recurrent patients maintained the original dose until they were not suitable for the medication.Results The 1 year recurrence rate in experimental group was 53.3％,that in control group was 86.6％ ( x2 =3.968,P ＜ 0.05).The 1 year survival rate in experimental group was 93.3％,that in the control group was 46.6％( x2 =7.777,P ＜ 0.05 ).The mean survival time of patients in experimental group was (24.6 ± 1.7 ) months (7 - 28 months),that in the control group was ( 16.4 ± 2.7 ) months ( 5 - 34 months ) ( x2 =7.154,P ＜ 0.05).Most adverse reactions in both groups were of grade Ⅰ - Ⅱ.The incidence of diarrhea and handfoot syndrome in experimental group is higher than that in control group.Conclusions Using Sorafenib for patients with primary hepatic carcinoma exceeding Milan criteria after liver transplantation may reduce carcinoma recurrence rate,and prolong patients' survival time.

Objective To explore the effects of ureteral stent on renal pelvic pressure and other urodynamic parameters. Methods Forty-one patients, 28 males and 13 females, with unilateral renal calculi and/or ureteral calculi were recruited in this study. The mean patient age was 47 years old (ranging from 20 to 72 years old). All cases were placed a 4.7 F ureteral stent and 16 F nephrostomy tube after minimal invasive pereutaneona nephrolithotomy (MPCNL). There was no hydronephrosis and residual crushed stone in the ureter after MPCNL in all cases. Renal pelvic pressure, intra-abdo minal pressure, detrusor pressure, bladder pressure changes during the filling and voiding phases with intravesical perfusion flow rate of 40 ml/min were recorded and analyzed. Results At the baseline, IPP0, IAP0, DP0 and BP0 were (33.1±17.0)cm H2O, (27.5±7.0)cm H2O, (3.3±2.9)cm H2O and (30. 9±7.2)cm H2O, respectively; At the maximum cystometric capacity during the filling phase, IPPvol, IAPvol Dpvol and Bpvol were (39.4±67. 3)cm H2O, (31.1±7.3)cm H2O, (10.7±6. 6) cm H2O and (41.6±10.3)cm H2O, respectively; At the maximum bladder pressure during the voiding phase, IPPmax, IAPmax Dpmax and Bpmax were (65.7±17.0)cm H2O, (33.7±9. 7)cm H2O, (41.9±7.8)cm H2O and (75.0±12. 8)cm H2O, respectively;There were statistical significance comparing between any of IPP0, IPPvol and IPPmax(P<0. 01). 27% (11/41)patients were with the pain in kidney area at voiding IPPmax (87.1±14.6) cm H2O, which was significantly higher than IPPmax (57.8±9.5)cm H2O of asyrnptomatic group (30 patients)(P<0. 01). In all cases, the renal pelvic pressure was higher than 40 cm H2O during the voiding phase. Conclusions Renal pelvic pressure increases during the filling phase after placing the ureteral stent, especially during the voiding phase. As renal function will be damaged by the high renal pelvic pressure, we should decrease the utilization of ureteral stent if possible. It is encouraged to remove the ureteral stent as early as possible.

BACKGROUND: It has been recently found that endothelial progenitor cells (EPCs) can promote injured endothelial healing. There is a supposition that in-stent restenosis possibly correlates with the number and/or activity of EPCs.OBJECTIVE: To comparatively observe the number and activity of circulating EPCs in patients with and without coronary in-stent restenosis, and to verify the above-mentioned supposition.DESIGN, TIME AND SETTING: This study, a comparative observation, was performed at the Department of Internal Medicine, Beijing Shijitan Hospital, Department of Internal Medicine, First Hospital, Peking University, and Department of Physiology and Pathophysiology, Peking University Health Science Center between March 2005 and May 2007.PARTICIPANTS: According to the coronary angiography, 15 patients were recruited into the restenosis group and 17patients with patent stents were selected into the control group.METHODS: Total peripheral mononuclear cells were isolated from blood of patients with restenosis or control subjects by Ficoll density-gradient centrifugation. These cells were plated on dishes coated with human fibronection. After 7 days in culture, the nature of adherent cells was confirmed by direct fluorescent staining with the use of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanide percholate-labelled acetylated low-dencity lipoprotein and fluorescein isothiocyanate-labeled ulex europaeus agglutinin-Ⅰ under a laser scanning confocal microscope. Cells demonstrating double-positive fluorescence were identified as differentiating EPCs.MAIN OUTCOME MEASURES: After 7 days of culture, EPCs were counted under an inverted microscope. Proliferation of EPCs was determined using the MTT colorimetric assay. Migration of EPCs was assayed using the scratch assay qualitatively. EPCs adhesion was performed by replating cells on fibronectin-coated dishes and then counting the adherent cells.RESULTS: The number of EPCs was significantly reduced in patients with in-stent restenosis compared with that in the control group (P = 0.001). The proliferative activities were impaired in the in-stent restenosis group than in the control group(P < 0.05). In addition, the migrative activities were also impaired in the in-stent restenosis group, but no significant difference in adherent activities existed between the two groups (P > 0.05).CONCLUSION: The number and functional activities of proliferation and migration of EPCs were decreased in patients with in-stent restenosis, which may be related to the number and/or activities of EPCs.

Humans ; Liver Cirrhosis, Biliary ; drug therapy ; Ursodeoxycholic Acid ; therapeutic use

Objective To investigate the effect of integrated traditional Chinese ( TCM ) and western medicine fast-track surgery ( FTS) in the perioperative management of laparoscopic cholecystectomy ( LC) .Methods One hundred and fifty patients with acute cholecysititis undergoing LC from June 2012 to January 2015, were recruited and divided randomly into 3 groups.Patients in group A (n=30) were treated with routine method in perioperative period, patients in group B ( n=60) were treated with western medicine FTS, and patients in group C (n=60) were treated with integrated TCM and western medicine FTS.The first exhaust time after operation, length of stay, times of clinic visit, symptoms, levels of IL-6, CRP and ALB, postoperative complications, readmission rate, reoperation rate, and patient satisfaction were evaluated in three groups.Results The first exhaust time after operation of group B and group C was earlier than that of group A [(25.16 ±8.36)h and (21.61 ±6.52)h vs.(36.06 ±10.88)h, P<0.05], and the first exhaust time of group C was earlier than that of group B (P<0.05).The length of stay of group B and group C were shorter than that of group A [(4.30 ±1.07)d and (3.98 ±1.16)d vs.(6.11 ±1.26)d, P<0.05].The nausea and vomiting, and abdominal distension of group C were lighter than those of group B [(0.27 ±0.08) vs.( 0.31 ±0.09); (0.35 ±0.09 ) vs.(0.40 ±0.13), respectively].There were no difference of the level of IL-6, CRP and ALB between group B and group C [(57.12 ±16.29) ng/L vs. (53.91 ±17.15) ng/L, (53.93 ±17.18) mg/L vs.(51.16 ±16.67) mg/L,(40.50 ±4.65) g/L vs. (41.01 ±4.60)g/L, respectively, all P<0.05].There was no difference among the three groups in the complication rate, readmission rate and reoperation rate(all P>0.05).Conclusion Integrated traditional Chinese and western medicine FTS in the perioperative period of LC can promote recovery, reduce symptoms and operation stress and maintain albumin level.

OBJECTIVETo study whether zearalenone (ZEA), a fungal estrogen, can transcriptionally up-regulate the expression of cytochrome 450 3A4 (CYP3A4) transcription by activating human steroid hormone and xenobiotic receptor (SXR).

METHODTransient cotransfection reporter gene assays were performed with human SXR expression plasmid and a reporter plasmid containing the SXR in the CYP3A4 gene promoter in HepG(2) cells.

RESULTSThe transcriptional induction of CYP3A4 by ZEA with a dose, time-dependent manner. ZEA at the concentrations of 0.01, 0.10, 1.00 and 10.00 micromol/L, respectively, could induce CYP3A4 with (1.50 +/- 0.21), (1.66 +/- 0.27), (3.04 +/- 0.82) and (3.96 +/- 1.16) folds, as compared with 0.1% DMSO. Results from a time-dependent study show that 1.00 and 10.00 micromol/L of ZEA for 12 to 48 hours could enhance the transcription of CYP3A4 with (3.69 +/- 1.34) and (5.18 +/- 1.50) folds, and 10.00 micromol/L of ZEA for 48 hours could induce the CYP3A4 gene expression (5.18 +/- 1.50) folds, as compared with 0.1% DMSO by activating human SXR.

CONCLUSIONZEA could induce the expression of the CYP3A4 gene transcription through activating SXR, possibly by affecting the other substrates of the CYP3A4, especially affecting the metabolism of drugs in the body.

Carcinoma, Hepatocellular ; pathology ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Genes, Reporter ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Transcription, Genetic ; drug effects ; Tumor Cells, Cultured ; Zearalenone ; pharmacology

AIM To study the anti-inflammatory and analgesic activities of ethanol extract of Toddalia asiatica Lamn.and the mechanism.METHODS Both inflammatory rat model induced by carrageenan and pain model induced by formalin were applied to investigating the analgesic effect of extract of Toddalia asiatica Lam.ELISA kit was used to detect the contents of β-EP,5-HT and PGE2 in serum of carrageenan-treated rats,contents of TNF-αand IL-1β in skin tissue of inflammatory rats,and content of LTB4 in serum of formalin-treated rats;immunohistochemical method was used to observe the SP and FOS protein expressions in rat spinal cord.RESULTS The ethanol extract of Toddalia asiatica Lam.could significantly reduce the rate of toe swelling.In the formalin test,the ethanol extract of Toddalia asiatica Lam.reduced not only the total licking time,but also the content of PGE2,especially in the high dose group.And lowered serum 5-HT contents were observed in all the three dose groups,but a much better performance was found in both the high and low dose groups,and the high dose group's capability in increasing serum β-EP content was also noticed.TNF-α and IL-1β contents in skin tissue were reduced in various dose groups.Middle and high dose groups inhibited FOS protein expression.And the content of LTB4 in serum was obviously decreased in the high dose group.CONCLUSION The anti-inflammatory and analgesic activities of ethanol extract of Toddalia asiatica Lam.may associate with its power in increasing β-EP in serum,decreasing PGE2,5-HT,LTB4 contents,reducing TNF-α,IL-1β contents in skin tissue,and lowering SP and FOS protein expressions in spinal cord.

Objective To study the effects of using 25 kDa linear and branched PEI to transfer plasmid DNA pEGFP -C1 (pDNA ,encoding the enhanced green fluorescent protein reporter gene ) to the basilar membrane of the C57BL/6 mice cochlea in vitro .Methods L -PEI/pDNA and B -PEI/pDNA polyplexes were generated in 0 .1M phosphate buffer solution (PBS) or 5% glucose solution .Polyplexes were characterized by transmission electron mi-croscopy .Agarose gel retardation assay was used to determine the plasmid binding ability of L -PEI and B -PEI . The toxicity was investigated by MTT assay .The transfection was firstly evaluated in 293T cell line ,and then the appropriate amount of PEI and plasmid were applied for cochlear explant transfection of P4 mice pups .Results Un-der the same condition ,B -PEI had better transfection efficiency than L -PEI ,but its toxicity was also higher . When generated in PBS ,the polyplexes had lower toxicity than in glucose solution .L -PEI-pDNA nanoparticles could transfect the spiral limbus fibrocytes ,some spiral ganglion neurons and supporting cells ,but the efficiency was low .Conclusion L -PEI could be used as the non -viral vector for the transfection of the cultured basilar mem-brane of P4 mice pups ,but it should be modified to reach higher efficiency .