Objective To set up an efficient and rapid method for detecting CK20 expression in peripheral blood of patients with colorectal carcinoma, and explore the relationship between CK20 expression and micrometastasis of colorectal carcinoma. Methods CK20 mRNA expression in peripheral blood of 36 patients with colorectal carcinomas was detected at various time points before and after operation by fluorescent RT-PCR. The mutation of p53 gene was also detected using SSCP. Results The positive rate of CK20 mRNA expression in peripheral blood before and 24 hours after operation was 75%(27/36) and 83.3%(30/36), respectively. After the short-term chemotherapy, the positive rate of CK20 mRNA expression was 36%(13/36). No mutation of p53 gene was found in those patients. Conclusion Fluorescent RT-PCR for detecting CK20 mRNA expression in peripheral blood was simple and efficacious. It was useful to monitor micrometastasis of colorectal carcinoma.

Objective To prepare the nanometer particles (NPs) of chitosan-5-fluorouracil (5-FU), and to study its characters of drug release in vivo. Methods Cross-linking polymer technique was employed to prepare the chitosan-5-FU NPs, and their characters were detected by scanning electron microscope and photon correlation spectroscope (PCS); the loaded capacity was measured by ultraviolet spectrophotometry; high-performance liquid chromatography was used to detect the characters of drug release in vivo. Results The chitosan-5-FU-NPs were found to be round or elliptic in shape, with 120 to 150nm in diameter and having a good dispersive ability. The loaded capacity was 31.000%?0.001%. The blood concentration of 5-FU was controlled at 76?g/L, and it was maintained for 48h. Conclusion As a vehicle, chitosan-5-FU NPs can change the behavior of pharmacokinetics and prolong the cycle time of 5-FU in vivo. Chitosan-5-FU NPs ran reduce the initial burst and prolong the releasing time.

Objective To discuss surgical techniques of laparoscopic treatment for intrahepatic stones.Methods A total of 57 cases of intrahepatic stones from March 2000 to March 2005 were studied. Under laparoscopic visualization,openings of left and right hepatic bile ducts were exposed through a longitudinal incision in the common hepatic duct to the bifurcation of left and right hepatic ducts.Stones in left and right hepatic bile ducts were evacuated with the use of a lithotomy forceps,a tiny basket,or pressure irrigation.Then openings of secondary hepatic ducts were exposed through the dilated primary ducts for the removal of stones in secondary hepatic ducts.Sometimes even openings of tertiary hepatic ducts could be seen for stone removal.When mud-or sand-like calculi were encountered,repeated irrigation of bile ducts was carried out.Results Conversions to open surgery were required in 3 cases(3/57,5.3%).The time of operation was 75～275 min(136?54 min).Residual stones were found in 49 cases(49/57,86.0%),which required postoperative choledochoscopy for 1~4 times to clear the intrahepatic stones.Complete clearance of all intrahepatic stones was achieved in 53 cases(53/57,93.0%).In 2 cases of calculi impacted in the lower common bile duct,laparoscopic electrohydraulic lithotripsy was performed for stone extraction.Postoperative bile leakage occurred in 5 cases(5/57,8.8%),and was cured with tube drainage.Follow-up observations for 0.5~5 years(2.3+1.5 years) in 43 cases showed excellent outcomes in 38 cases(88.4%),good outcomes in 3 cases(7.0%),and poor in 2(4.6%).Conclusions Intrahepatic stones can be removed laparoscopically through a longitudinal incision in the common hepatic duct to the bifurcation of left and right hepatic ducts for the exposure of primary and secondary hepatic ducts,or even tertiary hepatic ducts.

Objective To investigate the cytotoxicity of ADM-PBCA-NP on L-02 cells.Methods L-02 cells were cultured in vitro and the LDH activity of supernatant liquid of culture cells was examined.The toxicity of ADM-PBCA-NP,ADM and PBCA-NP to L-02 cells by the MTT assay was also determined,and the haemolysis function of PBCA-NP with different concentrations was detected.Results The cytotoxicity of ADM-PBCA-NP,ADM and blank PBCA nanoparticles to L-02 cells under the 10-6 mol/L concentration range was not cytototic(grade 1).LDH activity of supernatant liquid of culture cells showed no differences between the 3 groups.Conclusions Nanoparticles of ADM-NP and PBCA-NP in the 10-6 mol/L concentration range have no significant toxic effect on L-02 hepatic cells;and in a certain concentration range,the cell compatibility is excellent and will not lead to hemolysis.

Objective To evaluate laparoscopy assisted radio frequency ablation (LRFA) and percutaneous radiofrequency ablation ( PRFA ) for the treatment of hepatocellular carcinoma ( HCC ).Methods A total of 525 HCC patients were enrolled,including LRFA group of 78 cases with 106 tumor nodules,and PRFA group of 447 cases with 565 tumor nodules.Results ( 1 ) In LRFA group complete ablation rate was 97.17％ (103/106); in LRFA group complete ablation rate was 93.09％ (526/565)(x2 =2.523,P =0.112).(2)The 1,3,5 year overall survival rate was 96.15％,55.12％ and 38.46％in LRFA group and 93.73％,48.54％ and 31.54％ in PRFA group respectively ( x2 =0.699,1.151,1.447 ; P =0.403,0.283,0.229 ).The 1,3,5 year disease-free survival rate was 94.87％,43.58％ and 28.21％ in LRFA group and 93.73％,48.54％ and 31.54％ in PRFA group respectively (x2 =0.915,0.303,0.174; P=0.339,0.582,0.676).The average disease-free survival time was 22.25 months in LRFA group and 21.53 months in PRFA group respectively.(3)The serious complications was 0％ (0/78)in LRFA group and 1.34％ ( 6/447 ) in PRFA group respectively.( 4 ) Recurrence rate was 23.07％(18/78) in LRFA group,and 34.89％ (156/447) in PRFA group ( x2 =4.189,P =0.041 ).Conclusions The therapeutical effect of LRFA equals that of PRFA,while enjoying lower recurrence rate,no serious complications and higher treatment safety.

Objective The existing laryngoscope can not be used to observe the ultrastructure of cells.As a result,doctors always have to conduct biopsies on some patients who can not be diagnosed.The timeconsuming procedure of biopsy may be detrimental to the timely diagnosis and treatment.To solve this problem,we aimed to design the integrative confocal-microscopic hard laryngoscope system (ICMHL).Methods ICMHL system integrated the technique of existing laryngoscope and confocal microscopy,which allows doctors to observe the macrostructure and microstructure of lesions at the same time. Results With the effect of fluorescein sodium,ICMHL system could record the dynamic changes of cells at a rate of 30 fps,and the confocal-microscopic host machine could photograph images at a rate of 230 fps and diagnose invisible minute lesions.Conclusion ICMHL system provides a new,real-time approach for clinic biopsy.

Objective To prepare chitosan (CS)-compound Yizhihao-nanoparticles (NP) and to investigate its antibacterial activity. Methods CS NPs were formed by the incorporation of CS and Na3PO4. CS-compound Yizhihao NPs were prepared by ion-cross-linking. The particle sizes and surface charges of CS NPs were determined by Malvern Zetasizer 1000-HAS and atomic force microscope (AFM), respectively. The antibacterial acitivity of CS-compound Yizhihao-NPs was studied in vitro and compared with that of compound Yizhihao powder. Results Malvern Zetasizer 1000-HAS and AFM demonstrated that the diameter of CS-compound Yizhihao NPs was (137.00±14.28)nm and CS NPs had (16.90±1.32)mV positive surface charges. The minimal inhibitory concentrations (MIC) of CS-compound Yizhihao NPs on Staphylococcus aureus,Pneumococcus,β-hemolytic streptococcus, and Escherichia coli were 1:32,1:32,1:16,and 1:2,respectively. The minimal bactericidal concentrations (MBC) of CS-compound Yizhihao-NPs on Staphylococcus aureus,Pneumococcus,β-hemolytic streptococcus, and Escherichia coli were 1:16,1:16,1:8, and 1:2,respectively. The antibacterial efficacy of CS-compound Yizhihao-NPs to Staphylococcus aureus,Pneumococcus,and β-hemolytic streptococcus had been improved significantly (P<0.05). Conclusion CS-compound Yizhihao-nanoparticles have obvious antibacterial activity to the Staphylococcus aureus,Pneumococcus,and β-hemolytic streptococcus,which lays the experimental foundation for new preparation of traditional Chinese medicine in future research.

0.05).(2) The transfection rate and velocity could be increased by addition of extra-magnetic field.(3) Transfected-p53 obviously inhibited the growth of HepG2 at G1 phase;the growth inhibition rate of HepG2 was 46%.Conclusions(1) FFN is a good vector for gene transfection,it can successfully transfect the gene into target cells.(2) Transefected-p53 can effectively inhibit the growth of HepG2.This result can establish a foundation for animal experimental research of gene therapy of malignancy.

ObjectiveTo observe the e ff ect of NO precursor or/and NOS inhibitor on the survival of acute liver failure( ALF) rats.MethodsModel of ALF rat was established by resecting 90% of the rat liver and the effect of NO prec ursor or/and NOS inhibitor was observed.Resu ltsAdministration of NO precursor significantly improved the liver, lung, kidney and bowel function. The rats′ survival rate at 24 h, and 72 h increased significantly, whereas NOS inhibitor deteriorated fu nctions of important organs(P

Objective To investigate the effects of intrathecal morphine on cell-mediated immunity. Methods Forty male SD rats weighing 250-300 g were randomly divided into 5 groups ( n = 8 in each group) : sham-operated group (F); saline group (NS) and 3 morphine groups (M1, M2, M3). The animals were anesthetized with intraperitoneal chloral hydrate 300-350 mg?kg-1 . Microspinal catheter was inserted into the subarachnoid space at the lumbar region according to modified Yaksh technique. Correct implantation of the spinal catheter was confirmed by aspiration of CSF. In the morphine groups, after 5 days morphine was continuously infused through the spinal catheter at 2.5 (Ml), 5.0 (M2) and 10 ?g?h-1(M3) for 7 days. In NS group normal saline was continuously infused instead of morphine. On the 7th day 5% formalin 50 fd was injected into the plantar surface of left hindpaw. The number of flinches, lickings and total time of licking were recorded for 60 min. Pain intensity scoring (PIS) (0-3, 0 = no pain, 3 = severe pain) was used to assess the antinociceptive effect of intrathecal morphine. The animals were killed after evaluation of pain intensity. Body weight and spleen weight were measured. Spleen index (spleen weight/body weight) was calculated. T-lymphocyte function was evaluated based on Concanavalin-A (Con A) induced splenocyte proliferation. Modified lactic acid dehydrogenase (LDH) release assay was used to assess NK cell activity. Results The PIS scores were significantly lower in group M1 , M2 and M3 than in F and NS group. The spleen index, splenocyte proliferation induced by Con A and NK cell activity were significantly suppressed in the 3 morphine groups Conclusion Intrathecal morphine has significant antinociceptive effect and suppresses T-lymphocyte proliferation and NK cell activity in a dose-dependent manner