Intrahepatic cholangiocarcinoma is an uncommon malignant tumor,and its incidence has been increasing in the recent 30 years. Since patients have no specific clinical manifestations in early stage,the diagnosis of this disease is often very difficult,with a low rate of radical resection in late stage and poor prognosis.Therefore,as for patients with intrahepatic cholangiocarcinoma,early screening and diag-nosis is of vital importance.Imaging examination is an important method for the diagnosis of intrahepatic cholangiocarcinoma,and when com-bined with laboratory markers and pathological examination,it can increase diagnostic rate and reduce the rate of missed diagnosis.It is im-portant in clinical practice to select reasonable methods based on the patient′s actual condition.

Objective To detect the gene expression difference between gastric cancer tissue and lymph node metastasis,screen different express genes,and study its mechanism of metastasis and the relationship with biological behavior.Methods Using U133plus 2.0 gene chip technology,we detected the gene expression difference between gastric cancer tissue and epithelial cells of lymph node metastasis in five patients,and screened out the differentially expressed gene Twist-l.In vitro,the cell proliferation and apoptosis level were measured by using gene over-expression and gene knockout.Metastasis ability of carcinoma cells was detected by cell scratch assay.The expression changes of epithelial-mesenchymal transition (EMT)-associated protein (E-cadherin,Vimentin) in Twist-1 overexpression and gene knockout cells were determined by Western blot.Finally,we detected the expression of Twist-1 in gastric cancer tissue,and its correlation with TNM stage was analyzed.Results The expression of Twist-1 was higher in epithelial cells than in gastric cancer tissue (12.12±3.21 vs.2.07±0.71,P＜0.01).There was no correlation of the expression of Twist-1 with cell proliferation(absorbance of cell proliferation:negtive control 0.84±0.16,null vector control 0.74 ±0.06 and Twist-1 expression cell 0.71 ± 0.07) and apoptosis [cell apoptosis rate:negtive control (2.05±0.08)％,null vector control (4.31±0.07)％ and Twist-1 expression (3.95±0.09)％],but cell migration ability enhanced.In Twist-1 over-expression group,the level of E-cadherin was decreased,while vimentin increased.Conclusions Twist-1 gene changes might be correlated with the metastasis of gastric cancer by the way of EMT.

We prepared protoplasts from Salvia miltiorrhiza Bunge suspension culture cells. Then, the protoplasts' vitality and functions were tested by fluorescein diacetate staining method and Fluo-3/AM flourescent probe. The optimal condition of protoplast isolation was Cellulase R-10 1.5%, Pectinase Y-23 0.3%, Macerozyme R-10 0.5%, 40 r/min 12 h, 600 r/min 5 min, and the protoplasts yield was 1.1x10(6) cells/g FW, the vitality was more than 95% by using fluorescein diacetate staining method. It has been confirmed that calcium fluorescent probe Fluo-3/AM can be successfully loaded into protoplasts.
Aniline Compounds ; chemistry ; Cell Culture Techniques ; Cellulase ; chemistry ; Fluorescent Dyes ; chemistry ; Protoplasts ; chemistry ; Salvia miltiorrhiza ; growth & development ; Xanthenes ; chemistry

Objective To study dIe ESBLs and plasmid-mediated AmpC enzymes in E.Coli and Klebsiella spp. with CLSI ESBL-screening test positive,confirmation test negative but cefepime susceptible.Methods Antimierobial susceptibility testing were performed by Kirby-Bauer(K-B)method.The genes encoding ESBLs and plasmid-mediated AmpC enzymes were detected by PCR Transfer of ESBLs or plagmid-mediated AmpC resistance was studied by conjugation experiments.The homology of donor (E.coli),recipient(E.coli J53)and their transconjugants were analyzed by ERIC-PCR DNA fingerprints of E.coli and Klebsiella pneumoniae were analyzed by PFGE as recommended bv PulseNet protocoL Results Of 18 isolates from Huashan Hospital,11 were E.coli.6 were Klebsiella pneumoniae and 1 was Klebsiella oxytoca.Antimicrobial susceptibility testing indicated all of 18 isolates were positive on the CLSI ESBL screening test but negative on the confirmation test.and all of isolates were susceptible to cefepime(a zoneof-inhibition diameter of≥18 mm wag considered to indicate susceptible).PCR results indicated that 9 of the 11 E.coli isolates predued CMY-2 AmpC enzyme.TEM,SHV,CTX-M,PER,VEB or SFO type β-lactamages were not identified.Of 6 Klebsiella pneumoniae isolates.5 were DHA-1 AmpC-producing strains.4 of the 5 DHA-1 AmpC-producing strains were coexistence of broad-speetrumβ-lactamaae or extended-spectrumβ-lactamase.including two producing SHV-11 and two producing CTX-M-14 and SHV-62 type ESBL respectively.One Klebsiella oxytoca wag also DHA-1 AmpC producing strain.Conjugation experiments indicated that both ESBLs and AmpC enzymes could be transfefred from donor to recipient.PFGE indicated that the DNA fingerprints of K.pneumoniae were difierent but seven CMY-2 AmpC-producing E.coli isolates from general surgieal ward were similar.Concluslons The main mechanism of antibiotic resistance in CLSI ESBLs-screening test-positive but eefepime.susceptible E.coli and KIebsiellaspp.is production of plagmid-mediated AmpC enzymes.Some strains produce both AmpC enzyme and ESBLs.Such strains should be reported as resistant to cefepime.The results suggest that laboratories should routinely conduct research on the ESBLs and plnsmid.mediated AmpC enzymes in Enterobacteriaceae in order to report antimicrobial susceptibility testing results more correcdy.

Objective To investigate the release feature of dexamethasone sodium phosphate from thermosensitive in situ gels in vitro. Methods Rotation rheometer was used to measure the changes of viscosity with temperature. The membraneless model was applied in assessing corrosion behavior of gel using a thermostatic shaker (50 r/min) at an amplitude of 2.5 cm, taking phosphate buffered solution (pH 7.2) as releasing media. The release behavior was investigated by HPLC on a C18 reverse column DiamonsilTM (250 mm?4.6 mm, 5 ?m). The mobile phase consisted of triethylamine solution-methanol-acetonitrile (38∶28∶34), pumped at 1.0 ml/min, and the detection wavelength was set at 242 nm. Results When the temperature was near to the sol-gel transition temperature, the viscosity rose suddenly. Taking dexamethasone sodium phosphate (2 ml, pH 7.2) as media, the gel dissolution and drug release rate followed the zero order kinetics, and the cumulative gel dissolution (Q1) and cumulative drug release (Q2) equations were Q1=0.8238t (r=0.999, P

Objectives To determine the incidence of de novo neoplasms after liver transplantation and methods of prevention.Methods The clinical data of 605 patients after liver transplantation carried out from May 2000 to December 2010 in our centre were analysed.The patient's gender,age,primary disease and the clinical characteristics of post-transplantation de novo neoplasms were analysed.Results The incidence of de novo post-transplantation neoplasms was 1.80％.There was one case of post-transplantation lymphoproliferative disorders (PTLD),one case of pulmonary blastoma,one case of acute myelogenous leukemia,one case of bladder tumor,one case of gastric carcinoma,one case of sigmoid colon carcinoma,one case of carcinoma of rectum,one case of renal clear cell carcinoma and two cases of hepatocellular carcinoma.Conclusion There was a low incidence of de novo posttransplantation neoplasm in our centre than that reported in the West,probably because of variations in patient selection,immunosuppression regimens used,patients' age and different patient ethnic groups.Early diagnosis and treatment of de novo neplasm helped to improve prognosis.

The efficiency and safe range of Lipofectamine2000 (LF2000)/bcl-xl applied in human keratocytes, the optimal ratio of LF2000/bcl-xl and the bcl-xl gene expression in human keratocytes were investiaged. By using trypan-blue staining, the effects of LF2000 and bcl-xl on the survival rate of the cultured human keratocytes were measured respectively. By using semi-quantitative RT-PCR, the efficiency and the expression of LF2000-mediated bcl-xl transfection into keratocytes were examined. The results showed that the survival rate of human keratocytes had no signficant change in the presence of LF2000 (20 microg/ml) or bcl-xl (10 microg/ml) for 24 h. LF2000 could effectively mediate the transfection of exogenous gene bcl-xl into human keratocytes. The best transfection efficiency could be obtained when the ratio of bcl-xl/LF2000 was 1:8. One day after transfection, the positive cells for bcl-xl could be detectable, and the positive rate reached the peak-on the posttransfection day 3 (48.3%), then gradually decreased. Fifteen days after transfection, there were few positive cells. It was suggested that LF2000 could effectively transfer the exogenous gene bcl-xl into human keratocytes without obvious toxicity during a concentration range. LF2000/bcl-xl may be likely to play an important role in gene therapy of human keratocytes.
Cations/administration & dosage ; Cornea/cytology ; Gene Therapy ; Keratinocytes/cytology ; Keratinocytes/*metabolism ; Liposomes ; Transfection ; bcl-X Protein/biosynthesis ; bcl-X Protein/*genetics

Objective To retrospectively investigate the endoscopic characteristics and pathological features of colorectal polyps in over-aged patients (≥80 years). Methods The 1617 colonoscopies performed between January 2006 and December 2010 were enrolled in our retrospective analysis. The detection rate, size, location, form and pathological feature of polyps in 150 over-aged patients were investigated, and they were compared with those in 832 young patients (＜60 years) and 635 old patients (60-79 years) with colorectal polyps. Results The detection rate of polyps was 62.0% in over-aged group, and was significantly higher than in young group and old group (30.2% and 48.7%, respectively, χ2=56.58 and 8.64,both P＜0.001). The malignant transformation rate of ascending colon polyp was 5.4% in over-aged group, and was significantly higher than in young group (1.2%, χ2=4.90, P＜0.05), but there was no significant difference between over-aged group and old group (3.9%, χ2=0.36, P＞0.05). There were no statistical differences in canceration rate, polyp diameter, morphology and adenomatous polyp rate among the three groups. The malignant transformation rate was significantly increased in adenomatous polyps with diameter over 2 cm, but didn't reach statistical significance. Conclusions The polyp detection rate is noticeably higher in the over-aged than in the old and the middle-aged, and the malignant transformation probability is also increased. The colonoscopy indications in the high risk age groups should appropriately be broadened, they should receive regular intervals follow-up, and undergo polypectomy in time if necessary.

Objective To evaluate the treatment experience on primary retroperitoneal neoplasms involving large blood vessels.Methods In this study 82 patients with primary retropetropeal neoplasms were treated surgically from 1990 to 2010 and the clinical data of these patients including preoperative radiological examinations and surgery were retrospectively analyzed.Result The sensitivity and specificity of preoperative imaging examination for identifing large blood vessel involvement was 70％ and 73％respectively.Compared with those of no large blood vessel involvement,patients with large blood vessels involvement had a lower rate of tumor complete removal,more intraoperative blood loss and longer operation time( P ＜ 0.05).In group of large vascular involvement,patients undergoing complete tumor resection were all alive after 12 months,and none of those undergoing palliative resection survived more than 9 months.Conclusions Preoperative imaging allows precise evaluation on the status of large blood vessel involvement in patients of retroperitoneal tumors.Involving large vessels often predicts inability in total resection of retroperitoneal tumors.

Objective To evaluate the prevalence and risk factors of posttransplantation metabolic syndrome(PTMS)in liver transplant recipients.Methods According to NCEP-ATP Ⅲ(2001)standard,we assessed for the presence of PTMS in 102 transplant recipients which have lived ≥ 5 yesrs after liver transplantation.Age ≥ 50 years,the male sex,history of smoking,preoperative obesity,preoperative hypertension,preoperative diabetes,and preoperative hypertriglyceridemia were selected for evaluation.Predictors for PTMS were tested using Logistic regression analysis.Results Among 102 patients assessed,the prevalence of PTMS was 51.9％ compared with 29.3％ pretransplant level.Obesity(body mass index ＞ 28 kg/m2),hypertriglyceridemia(＞ 1.7 mmol/L),high-density lipoprotein cholesterol ＜ 1.0 mmol/L(men)or ＜ 1.3 mmol/L(women),hypertension,and diabetes were significantly more common in patients after transplantation.The independent factors associated with PTMS were older age,preoperative obesity and preoperative diabetes.Conclusions PTMS is highly prevalent among transplant recipients,older age(＞ 50 years),preoperative obesity and preoperative diabetes are among the risk factors for PTMS.