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 comprehend the status of Helicobacter pylori (Hp) infection in type 2 diabetes mellitus (T2DM) patients, and study the relationship between Hp and T2DM. Methods Seven hundred and sixty-eight in-hospital T2DM patients (T2DM group, including new diagnosed 194 patients and re-diagnosed 574 patients) and 172 sex, age matched health individuals(control group ) were involved in this study. 14C-urea breath test (14C-UBT) was proceeded in both groups to determine Hp infection, and infection rates were calculated. Results Total Hp infection rate in T2DM group was significantly higher than that in control group [56.0%(430/768) vs 37.8%(65/172) ,P< 0.01], delamination analysis according to sex also showed significant differences between T2DM group and control group(P < 0.01). The rate of Hp infection in new diagnosed patients and re-diagnosed patients, female patients and male patients had no significant difference (P > 0.05 ). The rate of Hp infection in different age and sex had no significant difference (P >0.05 ). Conclusions The rate of Hp infection in T2DM patients is higher than that in non-T2DM patients.Hp infection has relationship with T2DM, and has no relationship with age, sex and course of disease.

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.

Objective To investigate the association between CYP2C19 polymorphisms and efficacy of clopidogrel in Chinese Han patients with coronary atherosclerotic heart disease.Methods A total of 176 Chinese Han patients who underwent percutaneous coronary intervention and were treated with dual antiplatelet therapy with aspirin and clopidogrel were enrolled.Polymorphisms of CYP2C19 * 2,* 3 were measured by PCR-restriction fragment length polymorphism (PCR RFLP).The correlation between the genetic variants and cardiovascular events was analyzed.Results In the 176 patients,the allele frequencies of CYP2C19 * 2 and * 3 were 34.38％ and 5.97％ respectively,the percent of patients with poor metabolizer genotypes (* 2/* 2,* 2/* 3,* 3/* 3) was 14.20％.Among the 176 patients,11 patients reoccured acute coronary syndrome,1 patient suffered from stroke,and 4 patients had stent thrombosis.The incidence of cardiovascular events was 9.09 ％.Thecumulative incidence of cardiovascular events had no significant differences between poor metabolizer genotype carriers and extensive and intermediate metabolizer genotype carriers (16.00％ vs.7.95％,x2 =1.683,P=0.195).Conclusions The frequencies of CYP2C19 poor metabolizer genotypes are higher in Chinese Han population than in Caucasians,but the incidence of cardiovascular events in Chinese Han population is not increased.

Objective To compare the clinical effects of patellar resurfacing with patella reservation and patellaplasty in the total knee arthroplasty(TKA)for osteoarthritis so as to discuss appropriate management of patellas in TKA.Methods A retrospective study was conducted on 198 patients with osteoarthritis treated by TKA from January 2002 to December 2008.There were 62 patients managed by patellaplasty(patellaplasty group)and 136 patients by patellar resurfacing with patella reservation(patellar resurfacing group).The osteophytes of the patella were removed to make the articular surface of patellas similar to the primary one.Knee Society Score(KSS),Bristol patellar score,satisfaction survey and evaluation of joint range of motion(ROM)were performed during the regular follow-up.Incidence of postoperative anterior knee pain were analyzed and X-ray films were reviewed to understand the condition of the implants.Results A total of 125 patients were followed up,including 43 patients from the patellaplasty group and 82 from the patellar resurfacing group.The mean follow-up period was 51 months(range,36-80 months).Both groups showed significant improvement of each score postoperatively.No significant differences were found between the two groups regarding KSS score,patellar score and patient satisfaction in the follow-up one year later;but KSS function score in the patellar resurfacing group was superior to that in the patellaplasty group,with statistical difference.The incidence of anterior knee pain of the two groups had no statistical significance.Postoperative radiographs revealed no significant differences between the two groups concerning patellofemoral congruence,incidence of postoperative anterior knee pain or incidence of patellar related complications.Conclusion Patellar resurfacing with patella reservation for osteoarthritis in TKA can achieve satisfactory patellofemoral congruence and low incidence of intraoperative anterior knee pain,with comparable mid-term results with patellaplasty.

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.