pancreatic carcinoma by some signal transduction.

Objective To investigate nerve growth factor (β-NGF) and its receptors expression in human pancreatic ductal adenocarcinoma. Methods Expression and distribution of β-NGF, tyrosine kinase A (TrKA) and P75NGFR were detected in operation tissue specimens of pancreatic ductal adenocarcinoma with immunohistochemistry and real-time PCR. Relations of β-NGF and its receptors with clinicalpathological characters, especially nerve invasion were analyzed. Results β-NGF and TrKA expression are higher in pancreatic adenocarcinoma than normal pancreas, and the differences are significant (P < 0. 01). Β-NGF and TrKA expression are associated with the differentiation grades(DG), lymphatic node metastasis, nerve invasion and surgical pathological stages. Poorer of DG and later stages, more expression of β-NGF and TrKA. Β-NGF and TrKA expression have positive correlations. Β-NGF, TrKA and P75NGFR mRNA expression have significantly increased 3.84,4. 23 and 2. 41 times than normal tissues by real-time PCR, respectively. Conclusions β-NGF and TrKA might play potential rules in carcinogenesis for pancreatic cancer,have affinity with clinicopathological characters of pancreatic cancer. Β-NGF and TrKA may have mutual effect in signal transduction leading to perineural invasion of pancreatic carcinoma.

ObjectiveTo assess the satisfaction of general practical students' perception with the two modes of histologic laboratory instruction,and obtain the advantages and disadvantages of each mode.MethodsWeb-based database was established and in the questionnaires were delivered to the general practical students ( n=193 ).Results75％ of students agreed that the case study component of the webbased laboratory instruction was arranged in order.82％ of students identified with the detailed word explanation attached to every image presented.ConclusionStudents support the progression of web-based histologic laboratory instruction,but it is difficult to replace traditional microscope with web-based database in histologic laboratory instruction.

It was estimated that about 428 species of genus Corydalis are distributed all worldwide, with about 298, especially 10 groups and 219 species being uniquely spread in China. The genus Corydalis have been widely employed as folk medicines in China, especially as traditional Tibetan medicines, for treatment of fever, hepatitis, edema, gastritis, cholecystitis, hypertension and other diseases. The phytochemical studies revealed that isoquinoline alkaloids are its major bioactive ingredients. The extensive biological researches suggested its pharmacological activities and clinic applications against cardiovascular diseases and central nervous system, antibacterial activities, analgesic effects, anti-inflammatory, anti-oxidation and anti-injury for hepatocyte, and so on. As an effort in promoting the research of pharmacodynamic ingredients, this article presents an overview focusing on the distribution, phytochemical and pharmacological results of Corydalis species that have been applied in traditional Tibetan medicinal, hopefully to provide a reference for the new Tibetan medicine development from Corydalis plant resource.
Alkaloids ; chemistry ; pharmacology ; Animals ; Anti-Infective Agents ; chemistry ; pharmacology ; Corydalis ; chemistry ; classification ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Mice ; Molecular Structure ; Phytotherapy

Purpose: This study aimed to evaluate the effectiveness of an interactive virtual reality (VR) play intervention including instructional play and emotional catharsis play sessions in reducing children's pain and fear during intravenous placement. Methods: A randomized controlled trial with parallel groups was conducted. The sample consisted of 134 hospitalized children aged 6–12 years (intervention group: n = 69; comparison group: n = 65). The intervention involved one immersive intravenous scene in VR before the actual intravenous placement and one emotional catharsis VR play after injection. The comparison group received an educational photo book about intravenous placement before receiving intravenous placement. The children and their caregivers rated their pain and fear by using the Wong–Baker FACES Pain Rating Scale and the Children's Fear Scale. The time required for successful intravenous insertion was also compared between the two groups. Results: Children's pain (p = .028) and fear scores (p = .004) were significantly lower in the intervention group than in the comparison group. Their caregivers' pain and fear scores (both p < .001) were significantly lower in the intervention group. The time required for successful intravenous insertion did not differ significantly between the intervention and comparison groups. Conclusions The interactive play intervention with VR effectively reduced children's levels of pain and fear during the intravenous placement procedure. The results of this study can serve as a reference for the implementation of a feasible, child-friendly care practice for clinical intravenous placement in school-aged children.

OBJECTIVESecond mitochondria-derived activator of caspase (Smac) is a recently identified, novel pro-apoptotic molecule, which is released from mitochondria into the cytosol during apoptosis. Smac promotes activation of caspases by neutralizing members of the inhibitor of apoptosis proteins (IAPs) family, such as X-linked inhibitor of apoptosis protein (XIAP). The objective of the study was to examine the pro-apoptotic effect of human Smac gene on Burkitt's lymphoma Raji cells.

METHODSThe full length cDNA of human Smac gene was amplified by reverse transcription-PCR from total RNA of HEK-293 cells. The PCR product was ligated with linearized vector pGEM-T-easy supplied in the TA cloning kit and sequenced. The correct cDNA of full length Smac was subcloned into eukaryocytic expression vector pcDNA3.1/myc-his and transfected into human Burkitt's lymphoma cell line Raji by lipofectamine-mediated transfection. The expression of full length Smac was determined by Western blot. Morphological observation was done with the laser scanning confocal microscope by double staining the Raji cells with Hoechest 33,258 and propidium iodide. Flow cytometry was used to evaluate apoptosis. Relative caspase-3 activity was determined by colorimetric assay.

RESULTSRecombinant eukaryocytic expression vector pcDNA3.1/Smac, which contained full length Smac, was successfully constructed. After pcDNA 3.1/Smac was transfected into human Burkitt's lymphoma Raji cell line for 24 hours, Raji cells showed apparent apoptosis with a percentage of (43.7 +/- 2.5)%, which was higher than that of non-transfected group and free vector-transfected group (P < 0.05). Compared with non-transfected group (0.136 +/- 0.036) and free vector-transfected group (0.138 +/- 0.026), the relative caspase-3 activity of Raji cells transfected by pcDNA3.1/Smac (0.936 +/- 0.041) was significantly enhanced (P < 0.05).

CONCLUSIONTransfection and expression of human Smac gene could significantly induce apoptosis of human Burkitt's lymphoma Raji cells. The mechanism is associated with the increase of caspase-3 activity.

Apoptosis ; genetics ; Blotting, Western ; Burkitt Lymphoma ; genetics ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; DNA, Complementary ; Flow Cytometry ; Genetic Vectors ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Mitochondrial Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; methods

Objective A resistin binding peptide (RBP) was selected by phage display in our previous work. Studies had shown that RBP could antagonize the role of resistin on the lipid metabolism and endocrine function of adipose tissue, but whether RBP affects the insulin secretion of pancreatic cells is still unknown. The aim of this study is to assess the effect of RBP on basal insulin secretion in RINm5F insulinoma cells. Methods The cell viability was measured by 3-[4,5-dimethyhhiazol-2-yl]-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay. The supernatants were assayed for insulin content by enzyme linked immunosorbent assay (ELISA). Reverse transcriptase-PCR assay and Western blotting were used to determine the expression of glucose transporter 2 (GLUT2) involved in insulin secretion. Cytosolic Ca2+, the trigger of insulin exocytosis, was analyzed with the fluorescent probe FURA-3/AM. Results RBP did no effect on the cell viability with a concentration of 10-8-10-12mol/L of 2 hours intervention. But it stimulated basal insulin secretion of RINm5F cells, accompanied by up-regulated increased expression of GLUT2 and elevated concentration of cytosolic Ca2+. Conclusion RBP could stimulate basal insulin secretion without affecting the cell viability.

Objective To explore the treatment of cervical tracheoesophageal fistula (TEF) with complicated or remnant iaryngotracheal stenosis (LTS) and anterior neck defect (AND). Methods From 1980 to 2007,14 patients were diagnosed as TEF. Among them, 9 patients had complicated or remnant LTS, 3 patients had complicated AND, and 2 patients had TEF which were induced by Niekel-Titaium alloy mesh stent for treating benign esophageal stricture. All these patients were retrospectively studied in Tangdu Hospital. Treatment consisted of conservative therapy of TEF, staged surgical repair of TEF and laryngotracheal reconstruction according to the dimension (small or large) of TEF and complications. Results Four patients with small TEE (2-3 mm length) complicated LTS underwent laryngotracheal reconstruction stented with silicone T tube and TEF was adopted conservative treatment. The TEF and LTS were healed. Six patients with larger TEF (10-25 mm length) were repaired by staged surgical repair of TEF and laryngotracheal reconstruction. Among them, 3 eases had complicated LTS and AND, 2 cases had rement LTS and 1 ease had TEF without complication. Two patients had TEF and LTS, whose TEF healed before laryngntracheal reconstruction, the remnant LTS were reconstructed and healed. During the follow-up ranged from one to ten years, 12 patients were successfully treated without complications. One patient with TEF and LTS was treated only LTS because of a segment of esophagus was closed and treated with esophugngastrostomy in the department of thoracic surgery after LTS was successfully reconstructed and cured. One patient died of bleeding and asphyxia induced by the Nickel-Titanium alloy stentt because of the stent had not been taken out. Conclusion The small cervical TEF complicated or remnant LTS can be treated by laryngotracheal reconstruction and conservative treatment of TEF at the same time. A larger TEF complicated LTS should be treated by staged repair of TEF and LTS.

OBJECTIVETo explore the influence of different thawing temperatures on the morphology and type I collagen metabolism of the human fibroblasts processed at - 10 degrees C in vitro.

METHODSIn vitro cultured human fibroblasts were randomly divided into control, 20 degrees C thawing, and 37 degrees C thawing groups. After being frozen at -10 degrees C, the cells in the latter two groups were thawed at 20 degrees C and 37 degrees C, respectively. The cell proliferation was assessed with MTT method and was expressed by absorption under 570nm (A ), The morphological change of the cells was observed with inverted phase contrast microscope, the change in the intracellular content of collagen was determined with immunohistochemistry, and the extracellular content of collagen was assayed with ELISA.

RESULTSIn 20 degrees C thawing group, the absorbance decreased at first and increased thereafter, and they were obviously lower than that before freezing (0.95 +/- 0.16, P < 0.05 or 0.01). Cell dehydration and shrinking, cytoplasm loss and increased ratio of cytoplasm to nucleus were found in the survived fibroblasts. The cells proliferated actively at 72 and 96 hours after injury, with increased mitotic index and disordered arrangement. Compared with that before freezing (96.4 +/- 2.9) , the extracellular collagen content increased at first, decreased thereafter, and increased again slowly later (P < 0.05), while the intracellular collagen content decreased at first and increased thereafter (P < 0.05). The collagen metabolism in 37 degrees C thawing group was no difference compared with that in control group. Some cells undergone a floating period before adhering to the culture dish walls.

CONCLUSIONCell dehydration after low temperature treatment could protect the cells from damage. Proper thawing temperature could be beneficial to the cell resuscitation. Comparing with slow thawing, rapid thawing could minimize the cell damage.

Cells, Cultured ; Collagen Type I ; metabolism ; Cryopreservation ; Fibroblasts ; cytology ; metabolism ; Freezing ; Humans