【Objective】 To detect the copy numbers of brain-deriv ed neurotrophic factor (BDNF) gene in BDNF transfected PcDNA3.1(+)/BDNF/CHO cel ls with fluorescent quantitative polymerase chain reaction (FQ-PCR). 【Methods 】 BDNF DNA were amplified by GeneAmp 5700 Sequence Detection System with eq ual quantitative genomic DNA of PcDNA3.1(+)/BDNF/CHO, PcDNA3.1(+)/CHO and CHO cells as tamplates respectively. The process was repeated 30 times for every sam ples. The results were analyzed using q test. 【Results】 The copy numbers o f BDNF of PcDNA3.1(+)/BDNF/CHO cells and PcDNA 3.1(+)/CHO and CHO cells were 9 5 164±12, 31 622±10, 31 622±11 respectively. The copy numb ers of BDNF of PcDNA3.1(+)/BDNF/CHO cells were as three times as those of the P cDNA3.1(+)/CHO and CHO cells. The copy numbers of the two latters were the same . 【Conclusion】 The results clearly show that the PcDNA3.1(+)/BDNF/CHO cells h arbor two BDNF DNA copies.

With the rapid development of medical imaging technique, iodinated contrast media (IOCM) has become the most commonly used agent in performing imaging diagnosis and interventional therapy. The use of IOCM, especially the use of non-ionic iodine contrast media, has been swiftly increased in recent years. The accompanying untoward reactions, such as allergic reactions, cardiovascular reactions, contrast media nephropathy, thyroid toxicity, etc. prove to be the new problems that greatly perplex the medical circle. With the continuous improvement of the knowledge to the untoward reactions, more and more physicians have paid attention to contrast media induced thyroid toxicity. At present, researches concerning iodine contrast media thyroid toxicity are still few, and the relevant research is particularly rare in China. This paper aims to make a review about the influence of different kinds of iodine contrast media on thyroid function.

Objective: To investigate the influence of lidocaine on systemic inflammation in the perioperative ventricular septal defect (VSD). Methods: Twenty patients, scheduled for ventricular septal defect were randomly divided into 2 groups: lidocaine and control groups. Before rebeat lidocaine 1 mg/kg was given.The venous blood samples were obtained from the central venous at the following points: after induction of anesthesia and before cardiopulmonary bypass(CPB,T1),1 h after CPB(T2),2 h after CPB(T3), and 4 h after CPB(T4). IL-6 and IL-8 were determined by radio-immunoassay. Results: Compared with those at T1, the levels of white blood cells,polymorphonuclear neutrophils,IL-6 and IL-8 increased significantly from T2 to T4 in both groups. IL-6 and IL-8 levels reached the peak at T2. Compared with those in control groups, IL-6 level decreased obviously in lidocaine group from T2 to T4, but IL-8 level remained unchanged significantly. Conclusion: Under CPB and VSD repair the systemic inflammation is obvious, reaches the peak 30 min after CPB and persists to 4 h after CPB. Perioperative administration of lidocaine is effective against the inflammation.

Objective To investigate the role of signal transduction of toll-like receptors(TLRs)in immunological pathogenesis in children with acute idiopathic thrombocytopenic purpura(ITP).Methods Thirty children with actue ITP and 30 age-matched healthy children were studied.Real-time fluorescent quantitative PCR was used to evaluate the levels of TLR 1-10 and signal transducing molecules,and cytokines associated with TLRs,such as IL-1?,granulocyte-macrophage colony-stimulating factor(GM-CSF),macrophage colony-stimulating factor(M-CSF)and IFN-?/? mRNA.Expressions of co-stimulatory molecules such as CD40,CD80 and CD86 in mo-nocyte/macrophage(MC)was analyzed by flow cytometry.Results Compared with healthy control group,the mRNA levels of TLR3,7,8 and 9 in ITP group were significantly up-regulated(Pa0.05).Transcription le-vels of MyD88-dependent and-independent pathway molecules such as MD-2,MyD88,IRAK-4,TRAF6,TAK1,TRIF,TRAM,TBK-1 and IFN-? were significantly up-regulated in acute ITP(Pa0.05).Conclusion Aberrant activation of toll-like receptors signaling may be one of the initiating factors of immune dysfunction in children with acute ITP.

Objective To study the immunologic and pathologic features of an accelerated rejection model of renal allotransplantation in presensitized monkeys.Methods The accelerated rejection model of renal allotransplantation was established in presensitized monkeys,which received donor skin transplantation in advance(n=3).The changes of donor specific antibody(DSA)levels in the recipient monkeys before/after skin and kidney transplantation were measured.The kidney grafts were examined for routine pathology,antibody and complement depositions,various lymphocyte subsets infiltration by HE staining,immunofluorescence,or immunohistochemistry.Results All renal allografts in 3 presensitized monkeys developed accelerated rejection within 4 days.In 2 presentized monkeys,the levels of DSA and their mediated complement-dependent cytotoxicity(CDC)were significantly increased after skin transplantation,and further markedly elevated at the time of kidney graft rejection.In the rejected renal grafts,massive C3,C4,C5b-9 and IgG deposits with few lymphocytes infiltration were found.Typical pathologic changes included severe arterionecrosis,thrombosis,interstitial hemorrhage,and infiltration of neutrophils.In the rest one presentized monkey,the levels of DSA and CDC were only marginally increased,and the pathological changes of the rejected renal graft were characterized mainly by the injury of renal tubules.Conclusion Presensitization by donor skin transplantation could elevate the levels of DSA and CDC in recipient monkeys,which resulted in severe antibody-mediated acute humoral rejection in most of the following renal transplants.

Objective Pancreas perfusion is an essential step in human islet isolation.To develop the new methods for introductal canulation,collagenase infusion and to observe their effects on islets isolation.Methods A total of 17 pancreases were digested from March 2005 to April 2010.The pancreases were distended by three methods:the standard method (n =3),the one-cannula method (n =11) and three-cannula method (n =3).In the standard group,the pancreases were completely cut into half at the mid-body.Two catheters were inserted into the main duct:one directed toward the tail and the other to the head.In the one-cannula method group,a long tube was inserted into the duct at the head,advancing to the tail In the three-cannula method group,pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct:one at the head (the first catheter) and two at the mid-body,one toward the tail (the second catheter) and the other toward the head (the third catheter).The pancreases were digested by improved Ricordi technique.Ficoll continuous density/grads centrifuge method was performed to purify the islets.DTZ staining was adopted to identify islets and count islet equivalent (IEQ). AO/EB fluorescence examination was used to count active islet percentage.Static glucose stimulating test (SGS) in vitro was designed to estimate islet function and calculate SI.Results The distension volume of the threecannula method group was 1.24 rnl/g pancreas,and higher than the other groups (for the standard group:0.71 ml/g pancreas; for one-cannula method group:0.96 ml/g pancreas,P＜0.05).The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g,which was significantly more than that in the standard group (1914 IEQ/g pancreas,P＜0.05).The purity and viability of the islets were 74 ％/79.3 ％,75.6 ％/79.4 ％ and 78.3 ％/84.0 ％ respectively in the three groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).ConclusionThe three-cannula method improved collagenase infusion and the islet yields.

Objective To investigate the expression and clinical significance of intergrin linked kinase (ILK) in pancreatic carcinoma.Methods ILK protein was detected by immunohistochemistry and Western blotting in 60 cases of pancreatic carcinoma and 32 cases of normal pancreatic tissue,and the relationship with the clinicopathological characteristics were analyzed.Results Immunohistochemistry showed ILK was expressed in cytoplasm and membrane of pancreatic carcinoma cells and the positive rate was 65% (39/60),which was significantly higher than 18.75% (6/32) of normal pancreatic tissue(P ＜0.05 ).Western blotting showed the expression of ILK in pancreatic carcinoma tissue was 303933±195116,which was significantly higher than 144613±30074 of normal pancreatic tissue(P＜0.05 ).In pancreatic carcinoma,the expression of ILK was correlated with clinical stage and lymph metastasis(P＜0.05 ),but not correlated with tumor cell differentiation(P＞0.05 ).Conclusions ILK protein was highly expressed in pancreatic carcinoma tissue and it was correlated with the degree of malignancy.

Objectives To investigate the role of lymphangiogenesis in the perineural micrometastasis of pancreatic adenocarcinoma. Methods The clinical data of 30 pancreatic adenocarcinoma patients who were admitted from Sep. 2005 to Oct. 2006 for extended radical surgery were collected. The samples including pancreatic cancer, adjacent tissue, lower bile duct, pancreatic tail, the structure surrounding the SMA (peripancreatic nerve plexus) and lymph nodes were collected during operation. They were subjected to conventional pathological examination. The lymphatic capillaries weredetected by double immunohistochemical staining and the lymphatic vessel density ( LVD) was measured. Results Intra-pancreatic and/or peripancreatic neural invasion was observed in 25 patients (83. 3% ) , of which 20 were found to have both the peri-pancreatic and intra-pancreatic neural invasion. The other 5 only had the intrapancreatic neural fiber invasion and there was no single patient with peri-pancreatic neural fiber invasion only. Peri-neural invasion was not significantly associated with patients' age, gender, lymph node metastasis, tumor size and the location (P > 0.05) , but was obviously associated with JPS clinical staging ( P < 0. 05 ). The mean intratumoral LVD was (4.2 ±3.4) per field, which was significantly lower than (11.3 ±6.9) per field of adjacent tissue and (10.8 ±4.4)per field of normal pancreatic tissue(P<0.01). The mean intratumoral LVD between adjacent tissue and normal pancreatic tissue was not statistically different. Lymphatic vessel invasion was observed in non-malignant tissues in 18 patients, and there was a distribution correlation between lymphatic vessel invasion and extra-pancreatic neural plexus invasion (P<0.05). Conclusions The incidence of peri-neural invasion was high, peri-neural invasion was associated with JPS clinical staging and lymphatic vessel invasion, which suggested the possibility of the cancer spreading by peritumoral lymphangiogenesis route into the peri-SMA neural plexuses.

Objective To explore the role of splenectomy in the prevention and treatment of small-for-size liver in rat models, as well as its pathophysiologic mechanism in the development of a small-for-size syndrome (SFSS). Methods The models of sham-operation and 80 % partial hepatectomy (PH) were used in rats. In the experiment group splenectomy was performed following 80% PH. The concentrations of serum tumor necrosis factor (TNF-α), the content of NF-cB p65 in liver nuclear extracts, the expression of TNF-α, intercellular adhesion molecular (ICAM-1), and proliferating cell nuclear antigen (PCNA) transcripts, the activities of serum aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), total bilirubin (TB), albumin (Alb) cholinesterase (CHE), and liver myeloperoxidase (MPO) were analyzed. Portal venous pressures (PVP),incidence of SFSS,and one-wk survival rate were measured. Results In the control rats,The PVP was obviously elevated immediately after PH. The level of NF-κB p65 was obviously increased at the first h and peaked at about 3rd h postoperatively. The transcription of TNF-α and ICAM-1 and the release of serum TNF-α were significantly increased 3 h after PH. Capillary endothelial cells of the livers strongly expressed ICAM-1 24 h after PH. Splenectomy significantly reduced the PVP and the content of NF-κB p65 in the livers in concurrence with the expression of TNF-α and ICAM-1 gene as well as the activity of MPO at the corresponding time points after PH (P＜0. 05), while increased the expression of PCNA gene (P＜0. 05). Administration of splenectomy resulted in a statistically significant decrease in AST, ALT, LDH, TB, the incidence of SFSS and increase in one-wk survival rate (P ＜ 0.05 ). Conclusion Splenectomy alleviates liver injury and promotes liver regeneration in small-for-size liver rats by reducing portal vein perfusion and pressure,and suppressing NFκB activation and subsequent expression of proinflammatory mediators.

Adolescent ; Adult ; Aspergillosis ; complications ; microbiology ; Aspergillus ; isolation & purification ; Candida albicans ; isolation & purification ; Candidiasis ; complications ; microbiology ; Female ; Follow-Up Studies ; Granulomatosis with Polyangiitis ; complications ; microbiology ; pathology ; Humans ; Male ; Middle Aged ; Mucor ; isolation & purification ; Mucormycosis ; complications ; microbiology ; Nocardia ; isolation & purification ; Nocardia Infections ; complications ; microbiology ; Retrospective Studies ; Young Adult