Carcinoma, Hepatocellular ; therapy ; Catheter Ablation ; Chemoembolization, Therapeutic ; Combined Modality Therapy ; Humans ; Liver Neoplasms ; therapy ; Liver Transplantation

Carcinoma, Hepatocellular ; surgery ; Hepatectomy ; methods ; Humans ; Liver Failure ; diagnosis ; prevention & control ; therapy ; Liver Neoplasms ; surgery ; Postoperative Complications ; diagnosis ; prevention & control ; therapy

Switching Mechanism At 5′ end of the RNA Transcript(SMART)is a technology used in biology researching,So far,there is no review only about SMART technology.So,the aim is to investigate the research developments of principles,methods and applications of the SMART technology.Based on some researches and combined with the review of the related literature at home and abroad,it analyzed and evaluated the latest development of the research on SMART.As the applications of the SMART technology expand in many fields day after day,it has been proved that the SMART technology is a very useful and efficient skill to construct full length cDNA library.As more and more researchers know this technology,the advantages of the SMART technology become obvious,and meanwhile the disadvantages of the SMART technology also show up.That is to say,the technology needed to be improved.

ObjectiveTo observe and compare the efficacy of two types of microencapsulated rat islets xenotransplanted into diabetic mice. MethodsThe mice diabetic model made with injecting 3% Streptozotosin through tail vein. Four groups were assigned: control group, naked islet transplantation group, alginate-BaCl2 microencapsulated islet transplantation group, agarose-PSSa microencapsulated islet transplantation group.300 islets were transplanted under the renal envelope of each diabetic mice respectively. ResultsThere were no significant difference in mean level of the blood glucose before transplantation among four groups. One week after transplantation, the respective mean level of the blood glucose in four groups were (7.26±1.56) mmol/L in alginate-BaCl2 microencapsulated islet transplantation group, (7.14±1.04) mmol/L in agarose-PSSa microencapsulated islet transplantation group, (7.42±1.52) mmol/L in naked islet transplantation group and (22.54±1.24) mmol/L in control. There were significant difference between the two encapsulated islet groups and the other two groups. The survived period of the two encapsulated islet transplantation groups were longer than that of the other two groups. The survived period of the alginate-BaCl2 microencapsulated islet transplantation group was longer than that of the agarose-PSSa microencapsulated islet transplantation group (92 d vs 56 d),the same as the time of keeping nomal blood glucose level (76 d vs 41 d). ConclusionMicroencapsulated rat islets with this two materials can survive in diabetic mice with their biological activity, and the alginate-BaCl2 microcapsules are better than the agarose-PSSa microcapsules.

Animals ; Humans ; Liver ; growth & development ; Liver Diseases ; Receptors, Notch ; metabolism ; Signal Transduction

Objective:To value the application of two bacteria colony-PCR methods in the screening of phage antibody library. Methods:Five positive monoclonal bacterium were respectively suspended in either deionized water or 0.1% Triton X-100, and then boiled to be used as template in PCR. . The DNAs products of PCR were extracted and digested by two enzymes, and then determined by electrophoresis. Results:The inserted genes were detected in all the 5 clones after PCR and enzyme digestion .Conclusion:Bacteria colony-PCR can be used in screening positive recombinant colonies. The bacteria colony-PCR method with bacteria colonies suspended in deionized water is valuable in large scale positive recombinant bacterium screening.

ObjectiveTo study the protective effects of tumor necrosis factor-α(TNF-α) antibody on pancreatic encephalopathy in rats. MethodsSixty SD rats were randomly divided into the normal control group, acute necrotizing pancreatitis induction group and TNF-α antibody treated group. Acute hemorrhage necrotizing pancreatitis model in rats were induced by retrograde injection of 5% sodium taurocholate into the pancreatobiliary duct. Serum TNF-α was detected and animals were killed 12 h after drug administration. The changes of brain water contents, leucocyte accumulation and adhesion were measured, and pathological studies of pancreas and brain were performed. ResultsIn group of TNF-α antibody treated, serum TNF-α level decreased, brain water contents and leucocytes accumulation and adhension decreased significantly than that of acute necrotizing pancreatitis induction group (P<0.05). The histopathological change of pancrea was alleviative. ConclusionTNF-α antibody can alleviate the brain damage in acute hemorrhage necrotizing pancreatitis.

Objective To propose a new blood purification modality-hemodialysis with plasma- based dialysate (HD-PBD) plus high volume hemofiltration (HVHF) for patients with liver failure, and to evaluate the effect of this treatment on plasma cytokines.Methods Twelve patients with liver failure were included in this study.All patients received HD-PBD therapy in the first 6 hours,and then were treated with HVHF for 24 hours with the same filter (AV600).The levels of TNF-?,IL-1?, IL-6 and IL-8 in plasma before and after HD-PBD plus HVHF for 6 and 24 hours were examined respectively by ELISA,and changes of clinical parameters were observed at the same time point. Serum bilirubin,total bile acids (TBA),serum ammonia,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected before and after treatment.Arterial blood gas analysis and the concentration of electrolytes were monitored before and after treatment.Results (1)HD-PBD for 6 hours was more effective than HVHF for 24 hours in removal of serum bilirubin and TBA(P＜0.05). (2)Serum ammonia,BUN,Ser,arterial blood HCO_3~-,PCO_2,PO_2 and electrolytes did not show significant difference before and after HD-PBD (P＞0.05),but these parameters significantly changed before and after HVHF (P＜0.05).(3)The average level of serum bilirubin was sharply decreased after HVHF for 24 h following HD-PBD(P＜0.05).(4)After HD-PBD plus HVHF,there was a marked reduction of the plasma levels of TNF-?,IL-6 and IL-8.Conclusions HD-PBD plus HVHF,a newly proposed modality for patients with liver failure,can effectively decrease serum bilirubin,TBA,BUN,Scr,ammonia and cytokines,and adjust water-electrolyte as well as acid- alkali balance.It is a low-cost,safe,simple and convenient therapy.

OBJECTIVETo investigate the expressions of glypican-3 (GPC3) mRNA in hepatocellular carcinoma (HCC) tissues and peripheral blood cells (PBCs), and to determine the values of GPC3 mRNA in the diagnosis of HCC and HCC micrometastasis.

METHODSUsing semi-quantitative and nested reverse transcription polymerase chain reactions (RT-PCR), we detected the expressions of AFP and GPC3 genes in the tissues of 41 HCC, 41 paracancer and 52 non-HCC liver samples (41 far from HCC tissues and 11 normal liver tissues), and in the PBCs of 67 specimens from subjects.

RESULTSThe semi-quantitative RT-PCR displayed GPC3 mRNA was expressed in all samples of tissues and PBCs, and the relative intensities of its expressions in HCC, paracancer, non-HCC liver tissues were 78.9 +/- 35.5, 30.6 +/- 21.6, 23.8 +/- 15.5 respectively. The AFP mRNA expression values were 61.2 +/- 32.6, 31.5 +/- 23.6, and 21.2 +/- 15.9 respectively. The expression of each gene in HCC differed significantly from those in other two kinds of tissue samples (P < 0.01). The expressions of GPC3 mRNA and AFP mRNA, accounting for 80.5% and 63.4% in all the HCC tissues, were higher than their respective peak values in the tissues of non-HCC liver (+1.96s), but the expressions of at least one of the two genes was elevated in 92.7% of all the HCC tissues. There was a significant difference between combined detection of two genes and single AFP mRNA detection in HCC tissues (P < 0.01). Clinicopathologically, AFP mRNA was related with the grade of HCC and serum AFP, while GPC3 mRNA was related with not only the grade of HCC but also the invasion of HCC. The relative intensities of GPC3 mRNA expressions in PBCs of 67 specimens was 15.9 +/- 9.0, and GPC3 mRNA expressed in three kinds of tissue samples were all stronger than its counterparts in PBCs (P < 0.01). The GPC3 mRNA expression values in PBCs of the HCC group and the non-HCC group were respectively 16.1 +/- 8.3, 15.6 +/- 10.2, there was no significant difference between the two groups. Of the HCC metastasis group and the HCC non-metastasis group, the respective GPC3 mRNA expression values in PBCs were 16.0 +/- 9.0 and 16.3 +/- 7.7, there was also no significant difference between the two groups. The nested RT-PCR showed that the positive rates of AFP mRNA expressions in PBCs from the HCC group and the non-HCC group were 56.1% and 23.1%, and the difference between the two groups was significant (P = 0.011). The positive rates of AFP mRNA expressions in PBCs from the HCC metastasis group and the HCC non-metastasis group were 80.9% and 30.0%, and there was also a significant difference between the two groups (P = 0.002).

CONCLUSIONSAlthough GPC3 mRNA is expressed broadly, it still may serve as a potential tissue biomarker in the diagnosis of HCC. Detecting the expression of the two genes in the tissues will improve the screening and diagnosis of HCC. GPC3 is prevalently transcribed in the PBCs, but we have not found any relationship between the GPC3 expression in PBCs and the metastasis or recurrence of hepatocellular carcinoma, thus we can not identify HCC micrometastasis with GPC3 mRNA.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; pathology ; Female ; Glypicans ; biosynthesis ; genetics ; Humans ; Liver Neoplasms ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; alpha-Fetoproteins ; biosynthesis ; genetics

OBJECTIVETo study the cytotoxic activity against tumor cells and cytokines production of spleen cells induced in vitro by murine 4-1BBL gene transfected Hepa1-6.

METHODSThe eukaryotic expression vector pCDNA3.1(+)-m4-1BBL was transfected into murine hepatocellular carcinoma cell line Hepa1-6 by Liposomes. Then the transfected cells were selected in medium containing G418 (400 - 800 micro g/ml) and termed as Hepa1-6-m4-1BBL. The TCV-m4-1BBL was obtained by treating them with mitomycin (MMC). Cocultivation TCV with syngeneic murine spleen cells, then the lymphocytes were tested for cytotoxic activity against Hepa1-6-wt cells and the supernatants were harvested for detecting the cytokines (IL-2, TNF-alpha and GM-CSF).

RESULTSHepa1-6-m4-1BBL cells expressed 4-1BBL protein with highest cell surface level. The 4-1BBL mRNA could still be detected in the cells when cultured 48 h after treated with MMC (80 mg/L). Comparing with TCV-Hepa1-6, the tumor cell vaccine derived from Hepa1-6-m4-1BBL (TCV-m4-1BBL) could induce a more efficient cytotoxic activity of syngeneic murine lymphocyte against its parental tumor cell Hepa1-6 (P < 0.05), but not against non-parental tumor cell H22 and NIH3T3. Higher levels of IL-2, TNF-alpha and GM-CSF were released by the splencytes after stimulated by TCV-m4-1BBL.

CONCLUSIONSThese results suggest the expression of m4-1BBL by tumor cells is effective in inducing antitumor immune responses.

4-1BB Ligand ; Animals ; Cancer Vaccines ; immunology ; Female ; In Vitro Techniques ; Liver Neoplasms, Experimental ; immunology ; Mice ; Mice, Inbred C57BL ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Tumor Necrosis Factors ; genetics ; physiology