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

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 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

Objective To prepare thermosensitive in situ gel of ibuprofen ( IBU) inclusion complex with poloxamer and evaluate its release behavior in vitro and in vivo. Methods The IBU inclusion complexes were prepared by lyopyilization. The poloxamer 407 and 188 were added as a base for the preparation of gel. The release of inclusion complex gel was evaluated by membraneless models. The pharmacokinetics was evaluated after intramuscular injecting IBU solution or IBU inclusion complex gel to New Zealand white rabbits. The drug concentrations in the plasma were measured by HPLC. Results The loading amount of IBU was 10. 24% . The corrosion of gel and the release of IBU correspond to zero-order kinetics. Compared with IBU solution,the tmax and t1/2 were prolonged and Cmax was degraded while AUC was enhanced obviously. Conclusion The IBU inclusion complex gel is used as a good injection with sustained-release which can improve the analgesia and an- ti-inflammatory effect of ibuprofen. And our study builds up the foundation for the use of water-insoluble drug in thermosensitive in situ gel.

Objective To investigate the effect of a new type of retrograde reperfusion on the intraoperative internal environment and hemodynamics during classic orthotopic liver transplantation (OLT).Methods 20 patients underwent classic OLT using a new type of retrograde reperfusion in our center.Blood sampling was done at different parts or time points including:before blood venting via the portal vein (PV) of the donor liver,5 ml of blood was collected from the superior vena cava (PSVC),inferior vena cava (PIVC) and portal vein (PPV).During retrograde reperfusion through the inferior vena cava (IVC),5 mL of blood was collected when the volume of blood venting reached 5 ml (T1),100 ml (T2),and 200 ml (T3),respectively.The differences of data were compared after the blood samples were analyzed.In addition,the number instances of postreperfusion syndrome (PRS) were recorded.Results All operations were completed successfully,in which PRS occurred in 4 patients (20％).The most notable findings are the significant changes of nearly all data at T1,T2 and T3,including pH value,PvO2,SvO2,BEecf,HCO3-,Lac,K +,and Ca2 +,compared with PIVC (P ＜ 0.05) and a trend toward recovery in all the data.Yet their levels at T3 did not come back to the levels at PIVC (P ＜ 0.05).Besides,for pH value,Lac,K +,HCO3-and BEecf,there were no significant differences between PSVC,PIVC and PPV (P ＞ 0.05).Conclusions During classic OLT,the main factors leading to a disordered internal environment after recirculation stem from venous retum within the donor liver.This new type of retrograde perfusion can eliminate some of the harmful metabolites inside the donor liver in time and to some extent reduce internal environment disorders as well as drastic hemodynamic fluctuations after recirculation.

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 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 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.