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Objective To investigate the therapeutic effects of cFLIP-siRNA tail venous injection on acute necrotizing pancre.atitis (ANP) rats,and explore the related mechanism.Methods Three pairs of cFLIP siRNAs (cFLIP-siRNA) and negative control(siRNA-NC) were designed and synthesized.The most effective siRNA to inhibit the expression of cFLIP gene was selected for the experiment.Thirty SD rats were treated with retrograde pancreaticobiliary duct injection of 5％ taurocholic acid sodium salt solution to establish ANP rat model.They were randomly divided into ANP group,siRNA-NC group and cFLIP-siRNA group with l0 rats in each group using random number method.The same volume of normal saline,siRNA-NC and cFLIP-siRNA were injected into tail vein,respectively.The rats were sacrificed after 24 h and the blood samples were colelcted.The levels of serum amylase and endotoxin were measured.The pancreatic tissues were collected and routine pathological examination was performed.The expressions of cFLIP-S and caspase-8 protein in pancreatic tissues were detected by immunohistochemistry and Western blotting.Results The levels of serum amylase in ANP group,siRNA-NC group and cFLIP-siRNA group were (1 286 ± 209),(1 297 ± 305) and (552± 256)U/L,the level of serum endotoxin were (136± 32),(128± 56) and (46± 23)ng/L,respectively.The pancreatic pathological scores were (4.97 ± 1.16),(4.92 ± 2.03) and (1.67 ± 1.98).The expression level of cFLIP-S protein were (8.56 ± 2.72),(9.12 ± 2.45) and (3.82 ± 1.46),respectively,which were significantly lower in cFLIP-siRNA group than in ANP group and siRNA-NC group (P ＜ 0.05),and the difference was statistically significant (P ＜ 0.05).The expression level of caspase-8 protein in the three groups were (2.25 ± 1.24),(2.41 ± 1.14) and (5.56 ± 1.79),respectively,which were significantly higher in cFLIP-siRNA group than in ANP group and siRNA-NC group,and the difference was statistically significant (P ＜0.05).However,there was no significant difference between ANP group and siRNA-NC group.Conclusions Inhibition of cFLIP gene expression can reduce pancreatic injury in ANP rats.The possible mechanism was that the up-regulation of caspase-8 can inhibit pancreatic cell necrosis and promote apoptosis.
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Objective To investigate the level of the red blood cell (RBC) distribution width and other related indicators to predict the prognosis in patients with chronic heart failure elderly.Methods Retrospective analysis was performed in the patients with chronic heart failure elderly which died in our hospital (experimental group) and 100 patients were selected randomly that with heart failure elderly being improved by treatment (control group).The difference was compared in the level of the RBC distribution width (RDW),high-sensitivity C-reactive protein (hs-CRP),homocysteine (Hcy),D-dimer (DD),and amino-terminal pro-brain natriuretic peptide (NT-proBNP) in two groups.Results The level of RDW,hsCRP,Hcy,DD,and NT-proBNP in experimental group were significantly higher than that in control group (P ＜0.01).Conclusions Chronic heart failure was common clinical critically ill in the elderly.There were important clinical significance and important predictors to detect the levels of RDW,hs-CRP,Hcy,DD,and NT-proBNP in predicting prognosis and evaluating the effect of treatment.
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OBJECTIVE@#To explore the effect on proliferation and invasion of human papillary thyroid carcinoma K1 cells by application of small hairpin RNA (shRNA) silencing TFF3 gene expression.@*METHOD@#Using liposome transfection method, TFF3-shRNA targeting of TFF3 gene will be transient transfected to papillary thyroid carcinama K1 cells, inducing the corresponding gene silencing. The experiment set up blank control group (Con group), negative control group (ConNC group) and interference group (TFF3-shRNA group). The TFF3 protein and mRNA expression were evaluated by RT-PCR, Real time-PCR, immunocytochemistry and Western blot in K1 cells after TFF3-shRNA transfected. CCK-8 method and Scratch test were used to detect the change of proliferation ability and invasion ability respectively.@*RESULT@#(1) The recombinant plasmid Ca # HSH018037-4-HIVmU6 carrying TFF3-shRNA transfected K1 cells successfully. (2) RT-PCR and Real time-PCR detected the expression of TFF3 mRNA, which was 0.38 ± 0.11 times as many as the blank control group (P 0.05). (3) Western blot show that after TFF3 gene silence induced TFF3 protein expression levels have decreased 59.5% (P < 0.01), The difference was statistically significant compared with the blank control group. (4) Cell scratch detects K1 cell invasion ability. The invasion ability of K1 cells in interference group (TFF3-shRNA group) reduced. The scratch width significantly decreased 57.1% than blank control group (P < 0.01). (5) CCK-8 kit detect cell proliferation ability. K1 cells grow significantly slower in the interference group (TFF3-shRNA group) than the blank control group through the analysis of the growth curve (P < 0.01). In the interference group (TFF3-shRNA group) proliferation inhibition rate of K1 cells at 6 h, 12 h, 24 h and 36 h, 48 h are 16.6%, 26.6%, 33.6%, 33.8%, 35.0% respectively. Compared with negative control group, proliferation ability of K1 cell decreased significantly.@*CONCLUSION@#Silenced TFF3 gene can cause the degradation of mRNA, reduce the protein translation , and inhibit the invasion and proliferation ability of K1 cell.
Assuntos
Humanos , Carcinoma , Genética , Patologia , Carcinoma Papilar , Linhagem Celular Tumoral , Proliferação de Células , Peptídeos , Genética , Plasmídeos , Interferência de RNA , RNA Mensageiro , Genética , RNA Interferente Pequeno , Genética , Reação em Cadeia da Polimerase em Tempo Real , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide , Genética , Patologia , Transfecção , Fator Trefoil-3RESUMO
Objective To observe the morphological changes of pancreatic tissue of thoracic duct ligated rats in fine and ultrastructural levels, and to determine whether lymph block animal model can affect pancreatic islet amyloid polypeptide(PIAP)deposit in rat pancreas. Methods At the 6th month after the operation, some pancreatic tissue sections of 16-month-old experimental rats were embedded in paraffin wax and stained with HE and Congo red;immunohistochemical staining was performed on some frozen sections, which were then observed with light microscope;transmission electron microscope (TEM) specimen preparation and observation were performed on other samples. Results HE and Congo red stained sections showed that the pancreatic glandular lobule space was widened, with significant connective tissue hyperplasia, and fat accumulation when the islet was stained indistinctly or vermeil and tissue space was broadened. The sections with immunohistochemical staining displayed the pancreatic islet as well as the tissues around it were stained into dark brown being positive with PIAP antigen. TEM observation showed the pancreatic glandular interlobule space was widened, while blood vessels and enlarged lymphatic vessels were visible;within widened pancreatic islet interstitial space, a great quantity of lipid droplets and some collagen fibril structures could be seen.Conclusion The ligation of thoracic duct can contribute to pancreatic lymph draining block, lymphagiectasis, connective tissue space and interstitial space widening, fat accumulation, and PIAP deposit in rat pancreas. These structural changes may affect the function of pancreatic islets.
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Objective To investigate the expression of PMN Fas/FasL,Caspase-3 from peripheral blood in patients with SIRS and observe the effect of emodin on the PMN apoptosis and explore its mechanism.Methods The samples of peripheral blood were collected from 8 patients(all suffer from acute pancreatitis)with SIRS and 6 healthy volunteers.The circulating neutrophils were isolated and cultured.There were three groups of in our study:control group,SIRS group and emodin-treated group.The change of the PMN apoptosis and the expression Ieve]of Fas/FasL.Caspas-3 were observed.Results Compared with healthy volunteers.the percentage of PMN apoptosis significantly reduced in patients with S1RS(P＜0.05).After emodin was used,the percentage of PMN apoptosis in patients with SIRS increased(P＜0.05).Compared with healthy volunteers,the expression of Fas and Caspase-3 reduced in patients with SIRS(P＜0.05).Emodin could significantly induce the expression of Fas and Caspase-3(P＜0.05).After 24 hours of in vitro culture.the expression of FasL was not detected.Conclusion PMN from patients with SIRS shows profoundly delayed rates of apoptosis in vitro compared with PMN from healthy volunteers,and the decreased percentage of PMN apoptosis is associated with the reduced expression of Fas and Caspase-3.Emodin can significantly inhibit the delayed PMN apoptosis in patients with SIRS by inducing the expression of Fas and Caspase-3.
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BACKGROUND: The basic way for the releasing pattern of secretory granules of neurohypophysis is still not clear, neither is the extracellular normal transport route of neurohypophysis polypeptide hormones to enter the cerebrospinal fluid.OBJECTIVE: To provide the morphological evidences for the structural foundations of the releasing pattern and extraeellular normal transport route for secretory granules or polypeptide hormones of neurohypophysis by observing the structure of rats' neurohypophysis.DESIGN: Randomized controlled study.SETTING: Department of human anatomy in a university.MATERIALS: The experiment was completed in the Department of Human Anatomy of Hebei Medical University from May 2003 to January 2004. Twelve healthy clean grade adult male Sprague-Dawley rats with a body mass of about 300 g were supplied by the Experimental Animal Center of Hebei Province.METHODS: The 12 rats were randomly divided into 3 groups with 4 rats in each group. The neurohypophysises of each group were respectively observed with light microscope, transmission microscope and scanning electron microscope.MAIN OUTCOME MEASURES: The microstructure and ultrastructure of the neurohypophysis.RESULTS:All 12 rats entered the final analysis. In the coronary section of rat hypophysis, pars distalis, pars intermedia and pars nervosa were discernable under the light microscope. Under the transmission electron microscope, the neurohypophysis was composed of unmyelinated nerve fibers, pituicytes and connective tissue abound in blood capillaries. The endothelium of the blood capillary belonged to the fenestrated type(50 nm), separated from perivascular space by basement membrane. The intact secretory granules (100 -300 nm) coated with membrane existed not only in the endings of the unmyelinated nerve fibers but also occasionally in perivascular space. Under the scanning electron microscope, the pituitary capsule was composed of simple squamous epithelial cells and subepithelial connective tissue. Many irregular epithelial openings(2-5 μm) were observed among epithelial cells. Secretory granules were seen frequently near the epithelial opening.CONCLUSION: The releasing pattern of secretory granules or polypeptide hormones of neurohypophysis involves a whole-releasing pattern together with granular membrane. After released into perivascular space, they enter easily into cerebrospinal fluid via interspace of tissue and epithelial openings rather than into blood circulation through the walls of capillaries, and then into the cerebrospinal fluid.
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Objective To explore the structural foundation of transport pathway of pineal secretions from the pineal body to the cerebrospinal fluid of subarachnoid space. Methods The pineal capsule of the superficial pineal body of 1.5-month and 12-month old rats was observed under scanning electron microscope(SEM).Results Cribriform and trumpet-shaped epithelial stomata were scattered on the pineal capsule.Cribriform epithelial stomata were seen mostly in 1.5-month old rats.They were composed of many round pores which pierced through the full-thickness of the periphery of the capsule endothelium.The pores were dense and ranged from 200-500 nm in diameter;trumpet-shaped epithelial stomata were seen in both 1.5-month and 12-month old rats.Trumpet-shaped epithelial stomata were located among epithelial cells of the pineal capsule.They were round or elliptic in shape and ranged from 1-4 ?m in diameter.On the surface of the pineal capsule,many secretory granules were observed.They were spherule and 8001 000 nm in diameter.Conclusion The releasing pattern of secretory granules of the pineal body could include a releasing of the whole membrane;the pineal secretions may be transported directly from the pineal body to the cerebrospinal fluid of subarachnoid space through the epithelial stomata of the pineal capsule.