Carcinoma of gallbladder is the most frequently encountered malignancy of the biliary system. Early diagnosis is very difficult and the tumour resection rate is low,and the prognosis of carcinoma of gallbladder is very poor. With the increasingly widespread acceptance of laparoscopic cholecystectomy (LC),the number of cases of unsuspected gallbladder carcinoma (UGC) has increased. However,management of UGC is a difficult issue in the absence of established guidelines. High age and history of stones are the risk factors for gallbladder carcinoma and they are related to UGC. Surgical handling and pneumoperitoneum play an important role in the metastasis. The surgical approach used for cholecystectomy would not seem to influence the outcome in patients with UGC. The tumour stage is the most important prognostic factor. To understand UGC better, we review its clinical characteristics,investigation,prognosis and especially the recent advances in the diagnosis and treatment of this disease.

Objective To investigate the levels of platelet-derived growth factor-BB (PDGF-BB) and its role in the pathogenesis of the patients with hepatic diseases.Methods The levels of serum PDGF-BB in 30 chronic patients (hepatitis B,26; hepatitis C,4),24 patients with post-hepatitis cirrhosis,30 patients with primary hepatocellular carcinoma,and 20 normal controls were measured by ELISA.The levels of PDGF-B mRNA in peripheral blood mononuclear cells were measured by reverse transcription polymerase chain reaction (RT-PCR).Results The serum levels of PDGF-BB and the levels of PDGF-B mRNA in peripheral blood mononuclear cells were significantly higher in the patients with hepatic diseases (F=1774.40,1037.42,P

Objective:To establish a precise three-dimensional finite element model of temporomandibular joint.Methods: On the basis of images of Chinese Visible Human, the reverse engineering technology was applied to reconstruct the Computer Aided Design(CAD) model of temporomandibular joint.Afterwards, the model was established. Results:A three-dimensional finite element model consisting of 66 122 nodes and 212 704 elements of temporomandibular joint including cortical bone, cancellous bone, mandibular dental arch, masticatory muscles, articular cartilage and periodontal ligament was constructed. Conclusion:The finite element model is more efficient and more precise.

Objective To summarize some problems about the management of acute pancreatic pseuclocyst(PPC) by CT guided percutaneous drainage(PCD).Methods The recent domestic and foreign literatures were reviewed in order to explore the research advancement,such as indication,applied time,technique skill,complication and curative effect by CT guided PCD.Results This operation could be early performed in patients with acute PPC,preventing and decreasing the incidence of possible complications resulted from the traditional long-term observation and waiting.The technique skill of operation was easy with low incidence of complications and the effect was good.Combined use of somatostatin might shorten the treatment time.Some patients could be postoperatively managed in out-patient,thus the expense could be cut down.Conclusions CT guided PCD is a minimally invasive operation and is easy to perform with high effective rate and low incidence of complications and low cost.Reasonable selection of the indications and improvement of equipments and operation techniques may be helpful to improve the curative effect.The extended application of this operation is advised.

Objective To construct the eukaryotic expression vector of pcDNA3.1(+)/NOR1 and analyze the effect of NOR1 on the liver cancer cells. Methods NOR1cDNA was cloned into eukaryotic expression vector pcDNA3.1(+), recombinant eukaryotic expressing plasmid pcDNA3.1(+)/NOR1 was transiently introduced into human liver cancer cell line HepG2 mediated by cation iron lipofectamin.The biology effect of NOR1 on the liver cancer cells was analyzed through the MTT test, trypan blue exclusion assay and flowcytometric analysis. Results The eukaryotic expression vector of pcDNA3.1(+)/NOR1 was successfully constructed.The liver cancer cell growth rate was obviously slow after it was transfected by recombinant plasmid pcDNA3.1(+)/NOR1 and the cell cycle from G0/G1 to S distinctly prolong.Conclusions Recombinant plasmid pcDNA3.1(+)/NOR1 can express in HepG2 cells and affect the growth of HepG2 cells.

Objective To establish the quality standard of Fengshi Bitong Capsule.Methods Notopterygh Rhizoma,Angelicae Pubescentis,and Saposhnikoviae Radix were identified by thin layer chromatography(TLC).Aconitine limit test in this drug also carried out by TLC.The contents of Isoimperatorin in the Notopterygh Rhizoma were determined by high performance liquid chromatography(HPLC).Results The spots in the TLC were fairly clear and no interference was shown in the blank samples.The aconitine limited in conformity with the 《Chinese Pharmacopoeia》 2010.Isoimperatorin showed a good linear relationship within a range of 3.668 96-183.448 00 μg· mL-1(r=1).The average of recovery was 96.42%,RSD=2.64%,respectively.Conclusion The established methods are highly specific,stable and reproducible,which can be used as quality control of fengshi bitong capsule.

Objective To construct a recombinant eukaryotic expression plasmid of glycosylphosphatidyl inositol(GPI)-anchored bcr/abl and explore its expression at mRNA and protein level.Methods The gene fragment encoding bcr/abl was amplified by PCR using the plasmid containing the cDNA sequence of P210 as template and then inserted into a eukaryotic expression vector pBudCE4.1.The constructed recombinant plasmid pBudCE4.1-bcr/abl was identified by restriction analysis and DNA sequencing.Lymphocytes were isolated from human peripheral blood and their total RNA was extracted.The gene fragment encoding GPI was amplified by RT-PCR using the obtained RNA as template and was inserted into the constructed recombinant plasmid pBudCE4.1-bcr/abl in order to anchor GPI and bcr/abl.The constructed recombinant plasmid pBudCE4.1-bcr/abl-GPI was transfected into COS-7 cells,and the expressions of objective fragment were detected by RT-PCR and Western blotting.Results The results of restriction analysis,PCR and DNA sequencing proved that GPI-anchored bcr/abl fusion fragment was correctly inserted into vector pBudCE4.1.The expression of bcr/abl fusion gene and fusion protein were identified in transfected COS-7 cells and on their membrane.Conclusion The recombinant plasmid pBudCE4.1-bcr/abl-GPI was successfully constructed and expressed on the membrane of COS-7 cells,which found a basis of cell immunity with GPI-anchored bcr/abl fusion gene.

BACKGROUND:Scaffolds made of chitosan and its derivatives play an important role in cellmigration and axonal regeneration. Chitosan and its derivatives have good histocompatibility, which is easy to make stem cells grow on the surface, thereby having a more broad application prospect in the nerve tissue engineering.
OBJECTIVE:To fabricate a thermosensitive hydrogel scaffold using chitosan/hydroxypropyltrimethyl ammonium chloride chitosan/sodium glycerophosphate (CS/HACC/GP), which is suitable for cellgrowth, and then, to observe the growth and proliferation of bone marrow mesenchymal stem cells on the scaffold.
METHODS:Chitosan was modified using quaternary ammonium salt and confirmed by Fourier transform infrared spectroscopy. The chitosan and quaternary ammonium salt of chitosan was mixed at a ratio of 8:1 to successful y prepare stable CS/HACC/GP thermosensitive hydrogel scaffold. Then, the gel ing was observed, and biosafety test was conducted.
RESULTS AND CONCLUSION:Fourier transform infrared spectroscopy showed the characteristic peak of quaternary ammonium groups. Cytotoxicity test showed that rat bone marrow mesenchymal stem cells cultured in hydrogel extracts had no toxicity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test showed that hydrogel extracts exerted no significant effect on the increase in body weight, and the biological safety of the scaffold was good. Under the scanning electron microscopy, bone marrow-derived mesenchymal stem cells grew and proliferated normal y in the scaffold. The results confirmed that the CS/HACC/GP thermosensitive hydrogel scaffold was successful y prepared in the experiment, which is suitable for the growth and proliferation of bone marrow mesenchymal stem cells.

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

Objective To observed the expression of serum TNF-α and IL-10 in rats with severe acute pancreatitis (SAP) at different altitudes,and to explore the relationships between TNF-α and IL-10,the pathological changes of the pancreas,and the experimental basis for clinical diagnosis and treatment of SAP.Methods 72 specific pathogen free (SPF) Wistar male rats were divided randomly into three groups:1 500 meters altitude (group L),3 300 meters altitude (group M),and 4 300 meters altitude (group H).These three groups were then each divided randomly into four subgroups:control (group n),6 hours after pancreatitis (group p 6 h),12 hours after pancreatitis (group p 12 h),and 24 hours after pancreatitis (group p 24 h).Pancreatitis was induced by intraductal administration of 5％ sodium taurocholate hydrate (NaTc).The rats were killed at 6,12,and 24 hours after NaTc injection in groups p.The group n rats were killed after 6 hours of pancreas observation.Blood samples and pancreatic tissues were collected post mortem and enzyme-linked immunosorbent assay (ELISA) measured serum TNF-α and IL-10.Results Compared with the control (group n),histopathological scores,IL-10,and TNF-α in the same altitude had a significant difference (P ＜ 0.05) in group p at each time point.In the same altitude of group p,histopathological scores and IL-10 were increased with time elapsed (P ＜ 0.05),while TNF-α was decreased with time elapsed (P ＜ 0.05).There was a significant difference between group Mp and Lp in histopathological scores,IL-10,and TNF-α (P ＜ 0.05),and the same result between group Hp and Lp (P ＜ 0.05),but there was no significant difference between group Hp and Mp (P ＜ 0.05).Meanwhile,IL-10 had a positive relationship with histopathological score,but TNF-α had a negative relationship with histopathological score.Conclusions The level of TNF-α increased with increasing altitude but significantly reduced with elapsed time.The level of IL-10 increased with both increasing altitude elapsed time.These results suggested that TNF-o and IL-10 might play a important role at different times in severe acute pancreatitis.