Aim To establish a gefitinib-resistant NSCLC cell line, and observe its pharmacological properties. Methods HCC827 cells were treated with hepatocyte growth factor( HGF) and increasing concen-tration of gefitinib to induce cell resistance. MTT assay was used to test cell viability and chemosensitivity. Clone formation and Edu staining were used to verify the inhibitory effect of gefitinib on cell growth. qRT-PCR and Western blot were used to assay the expres-sion of c-Met. Results The use of HGF greatly short-ened the induction time of HCC827GR resistant cells. gefitinib could significantly suppress cell viability, col-ony formation and cell proliferation of HCC827 , but had a weaker inhibitory effect on HCC827GR cells. HCC827GR overexpressed c-Met protein, and was highly sensitive to c-Met inhibitors. Conclusion The gefitinib-resistant HCC827 GR cells were induced suc-cessfully and efficiently. The growth of HCC827GR is dependent on the activity of c-Met protein, and it can be used as a phenotypic screening model of c-Met in-hibitors.

Objective To study the results of laparotomy and Cool-tip radiofrequency ablation to treat large liver tumors.Methods Laparotomy and Cool-tip radiofrequency ablation were carried out on 64 patients with large hepatic cancer.To destroy the tumor completely,for tumors of 3.0～4.0 cm in diameter,7 ablations were required; for 4.0～5.0 cm in diameter 15 ablations; for 5.0～6.0 cm in diameter 19 ablations; for 6.0～7.0 cm in diameter 40 ablations.Result The complete necrosis rate of laparotomy and radiofrequency ablation was 93.75％ (60/64).The short-term results were good.Conclusions Laparotomy and Cool-tip multipoint overlapping radiofrequency ablation for large liver tumors (tumor diameter＞3 cm) could result in a high complete necrosis rate and a low complication rate.It is a good radical treatment for unresectable and large liver cancer.

Objective To investigate the effects of improved end-to-end invagination pancreaticojejunostomy on the occurrence of pancreatic fistula after pancreaticoduodenectomy.Methods The clinical data of 396 patients who received pancreaticoduodenectomy at the Cancer Hospital of Shandong Province from January 2001 to January 2011 were retrospectively analyzed.All patients were divided into the improved group(235 patients)and traditional group(161 patients)according to different anastomotic methods.All the operations were done by the same surgical group,and the digestive tract was reconstructed by the Child method.Patients in the improved group received improved end-to-end invagination pancreaticojejunostomy,and patients in the traditional group received traditional end-to-end anastomosis.The volume of operative bleeding,operation time,incidence of pancreatic fistula and duration of hospital stay of the 2 groups were compared.All data were analyzed using the t test,chisquare test or Fisher exact probability.Results The operative blood loss,operation time and duration of hospital stay were(383 ±56)ml,(7.2 ± 1.0)hours,(21 ±3)days in the improved group,and(381 ±39)ml,(7.0 ± 0.5)hours,(22 ± 5)days in the traditional group,with no significant difference between the 2 groups(t =0.388,1.680,-1.835,P ＞ 0.05).No operative death was detected in the 2 groups,and the overall incidence of pancreatic fistula was 7.6％(30/396).The incidence of pancreatic fistula of the improved group was 0(0/235),which was significantly lower than 18.6％(30/161)of the traditional group(P ＜ 0.05).Patients complicated with pancreatic fistula in the traditional group were cured by drainage,somatostatin administration and parenteral nutrition.Conclusion Improved end-to-end invagination pancreaticojejunostomy can significantly reduce the incidence of pancreatic fistula after pancreaticoduodenectomy.

Adenocarcinoma ; metabolism ; pathology ; surgery ; ultrastructure ; Aged ; Carcinoembryonic Antigen ; metabolism ; Carcinoma, Neuroendocrine ; metabolism ; pathology ; surgery ; ultrastructure ; Cardia ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Microscopy, Electron, Transmission ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; ultrastructure ; Synaptophysin ; metabolism

Phylogenetic methods have been widely used to detect the evolution of influenza viruses.However, previous phylogenetic studies of influenza viruses do not make full use of the genetic information at the protein level and therefore cannot distinguish the subtle differences among viral genes. Proteotyping is a new approach to study influenza virus evolution. It aimed at mining the potential genetic information of the viral gene at the protein level by visualizing unique amino acid signatures (proteotypes). Neuraminidase gene fragments of some H5N1 avian influenza viruses were used as an example to illustrate how the proteotyping method worked. Bayesian analysis confirmed that the NA gene tree was mainly divided into three lineages. The NA proteotype analysis further suggested there might be multiple proteotypes within these three lineages and even within single genotypes. At the same time, some proteotypes might even involve more than one genotype. In particular, it also discovered some amino acids of viruses of some genotypes might co-reassort. All these results proved this approach could provide additional information in contrast to results from standard phylogenetic tree analysis.

Objective To investigate the changes of cardiac sinus node connexin 45 (Cx45), connexin 31.9(Cx31.9) of gap junction in rabbits'' sinus bradycardia model caused by dexmedetomidine, and to discuss whether the negative frequency and negative conduction caused by Dexmedetomidine are related to the expression changes of connexin.Methods Forty-eight healthy adult New Zealand rabbits were divided into three groups (n=16).Sinus bradycardia rabbits were prepared by intravenous injecting Dexmedetomidine through ear vein.After rabbits were anesthetized by sodium pentobarbital, basic procedures were quickly completed in order to monitor MAP and ECG.Rabbits in group C were injected with normal saline.Rabbits in group D1 were injected a loading dose of dexmedetomidine 10 μg/kg for 10 min, and then continuously pumped 5 μg·kg-1·h-1 for 50 min.Rabbits in group D2 were pumped a loading dose of dexmedetomidine 60 μg/kg for 10 min, and then continuously pumped 30 μg·kg-1·h-1 for 50 min.After observation hearts were quickly removed and sinus node tissue was dissected.The average optical density of sinus node Cx45, Cx31.9 were detected by immunohistochemistry, and the genes expression were detected by real-time quantitative.Results Cx45 gene expression of group D2 showed remarkable increase than groups C and D1(P<0.05), there were no significant differences between groups D1 and C, Cx45 average optical density change were consistent with the gene expression.Cx31.9 gene expression of group D1 showed a more remarkable increase than group C(P<0.05), there were no significant differences between groups D2 and C,D1 and D2, Cx31.9 average optical density changes were consistent with the gene expression.Conclusion Dexmedetomidine increases the expression of low electrical conductivity Cx45, Cx31.9 of gap junction on rabbits'' sinus node, which is one of the possibly reasons that slow down cardiac conduction velocity in sinus node.

BACKGROUND:Simulative dynamics provides advantages of repeatable and non-invasive to a model. Additional y, structural model of individualism improves the reliability of Finite Elemental Analysis. It is an optimal attempt to analyze mechanics of knee joint after virtual replacement surgery.
OBJECTIVE:To achieve dynamic information of contact stress upon knee joint surface by finite element analysis surgery for total knee arthroplasty postoperatively, and to provide objective data for further“surgery plan”.
METHODS:After scanning affected knee joint by CT/MR and scanning knee prosthesis by laser instrument, a model composed of prosthesis, knee joint as wel as its ligament was rebuilt computational y;dynamic lines were measured. After prosthesis instal ation&osteotomy performed by facility of Simulation in Mimics according to knee joint replacement standard, this model was imported into ANSYS so as for Meshing, Material assignment, load applied. Stress distribution was analyzed by Finite Element Method.
RESULTS AND CONCLUSION:The best finite element model of postoperative TKA 3D knee joint was obtained;dynamic data were tested to be approximately agreeable to those previous studies of direct measurement upon prosthesis. The experiment of analyzing structural deformation, stress distribution&internal energy change benefits to search the best position for prosthesis instal ation, osteotomy and surgical result prediction. Thus, these are indispensable data in surgery plan.

To construct a scFv library by phage display technique from the spleen cells of mice immunized with B3HM cells. Three mice were immunized with B3HM cells, and their spleen cells were harvested. The genes of VH and Vk were amplified by RT-PCR from the cDNA of the immunized spleen cells and a scFv-phage display antibody library was constructed. The capacity of library was measured,and the variety of the library was analyzed by digesting with restriction endonuclease BstNI.ScFv phage clones were randomly picked and identified phage-scFv clone by binding B3HM cells using immunofluorescein.A scFv library containing 5?106 individual clones which showed different patterns after digested with restriction endonuclease BstNI was produced. Individnal phage-scFv clone showed B3HM cells positive using immunofluorescein. A scFv library of anti-B3HM cell surface molecules has been constructed. It will be useful for finding out some novel genes of causing leukemia, and establishs the infarctate foundation of clarifying the pathogenesis of leukemiagenesis.

OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.

METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.

RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.

CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.

Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea

OBJECTIVETo investigate the changes in the expressions of p53, DKK (the inhibitor of Wnt pathway) and phosphorylated tau in rat bilateral hippocampus after beta-amyloid peptide (beta-AP) (25-35) injection, and observe the effect of saponin B from Anemarrhena asphodeloides Bunge (SAaB) in this model.

METHODSAfter bilateral injection of beta-AP (25-35) into the hippocampus of rats, RT-PCR was performed for observing the changes in p53 and DKK mRNA expressions and immunochemistry carried out to detect the changes in phosphorylated tau protein.

RESULTSRT-PCR showed increased p53 and DKK mRNA expression and immunochemistry revealed increased phosphorylated tau-positive cells in rat hippocampus after beta-AP (25-35) injection, and administration of SAaB significantly ameliorated these changes.

CONCLUSIONSAaB can significantly ameliorate beta-AP-induced tau hyperphosphorylation by inhibiting increased p53 and DKK mRNA expressions in response to beta-AP injection.

Amyloid beta-Peptides ; pharmacology ; Anemarrhena ; chemistry ; Animals ; Hippocampus ; drug effects ; metabolism ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Male ; Peptide Fragments ; biosynthesis ; genetics ; pharmacology ; Phosphorylation ; drug effects ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Saponins ; pharmacology ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; tau Proteins ; metabolism