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Objective:To explore the expression of miR-216a in patients with acute pancreatitis (AP) associated with acute lung injury (ALI) and its influence on endothelial cells permeability.Methods:40 AP patients admitted in Department of Gastroenterology of the First Affiliated Hospital of Navy Medical University from December 2015 to March 2016 were collected and were classified into AP with ALI (AP-ALI group, n=13) and AP without ALI (AP group, n=27) according to the presence or absence of ALI. 8 normal volunteers were enrolled in the control group. Blood samples were collected and the plasma samples were separated. Plasma RNA was extracted. miR-216a level in plasma was detected by RT-PCR. Plasma exosomes were extracted by exosome extraction kit and identified by the electron microscopy. Exosome RNA was extracted. miR-216a level in exosome was detected by RT-PCR. Plasma exosomes of AP-ALI patients were co-cultured with human umbilical vein endothelial cells (HUVEC, AP-ALI-HUVEC group) and anti-miR-216a transfected HUVECs (AP-ALI-anti-miR-216a HUVEC group) for 24 hours, respectively, and untreated HUVECs served as control group. Trans-endothelium electrical resistance (TEER) was measured by Millicell Ers-2 epithelial volt-ohmmeter to evaluate the cell permeability. Results:RT-PCR results showed that the expression level of plasma miR-216a in AP-ALI group (14.45±1.64) was significantly higher than that in AP group (11.08±1.6) and the control group (5.37±1.54) ( P<0.01). Under electron microscope, plasma exosomes were goblet like vacuoles, with the size of about 50-90 nm. The plasma exosomal miR-216a level in the AP-ALI group (14.03±1.58) was significantly higher than that in the AP group (10.86±1.31) and the control group (5.01±0.79), and the difference was statistically significant ( P<0.01). The resistance value of HUVEC in the control group was referred as 1, and the resistance ratio of HUVEC in AP-ALI-HUVEC group was 0.74±0.04, which was significantly lower than that of HUVEC in AP-ALI-anti-miR-216a HUVEC group (1.02±0.08), the difference was statistically significant ( P<0.01). Conclusions:miR-216a was highly expressed in plasma exosomes of AP patients with ALI. miR-216a can increase endothelial cell permeability, which may be associated with ALI during AP.
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Objective To investigate the early predictive value of several commonly used biochemical markers for predicting persistent organ failure ( POF ) in patients with hyperlipidemic acute pancreatitis ( HLAP) . Methods Clinical data of 157 patients with HLAP within 72 hours after the onset of first attack who were admitted to the Dept. of Gastroenterology in Changhai Hospital from January 2015 to December 2017 were retrospectively analyzed, including 106 cases without POF ( non POF group ) and 51 cases with POF ( POF group) . Hct, BUN, Cr, APACHEⅡand BISAP were recorded within 24 hours after admission. Receiver-operating characteristic ( ROC) curve was drawn to calculate area under the ROC curve ( AUC) and evaluate the performance of Hct, BUN, Cr, APACHEⅡand BISAP scores in predicting HLAP complicated with POF, which was compared by DeLong test. Results Values of BUN, Cr, APACHEⅡand BISAP were significantly higher in HLAP patients with POF than those without POF [(10. 30 ± 7. 43) vs (5. 34 ± 2. 26) mmol/L, (165. 31 ± 123. 93) vs (65. 61 ± 20. 82)μmol/L, (10. 22 ± 6. 22) vs (4. 61 ± 2. 99) points, (2. 61 ± 0. 87) vs (1. 42 ± 1.07) points], and the differences were all statistically significant (all P<0.05), whereas Hct was not significantly different between the two groups. The AUC of Cr and BUN for predicting POF was 0. 77(95％ CI, 0. 69-0. 86) and 0. 71 (95％ CI, 0. 61-0. 81), respectively, and the optimum predictive Cut-off values were 130 μmol/L and 8. 95 mmol/L, respectively. The sensitivity was 53％, and the specificity was 99％ and 94％;the accuracy was 84％ and 81％;negative predictive value was 81％, and positive predictive value was 96％ and 82％. DeLong test showed that predictive performance of BUN and Cr was not statistically different from that of APACHEⅡand BISAP. Conclusions Cr≥130 μmol/L and BUN≥8. 95 mmol/L can be used clinically to predict the presence of POF in HLAP, and the predictive efficacy were comparable to APACHEⅡand BISAP.
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Hematoma enlargement occurs in about 20％ ～ 30％ of patients with intraeerebral hemorrhage,leading to worsening of the medical conditions and severe economic and social burden.It is pivotal to explore its mechanism and predictors,establish reliable prediction model,to understand the occurrence,development and treatment of the disease.In this paper,the physiopathologic mechanisms,imaging predictors,biochemical predictors,prediction models of hematoma enlargement are summarized as follows.
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Objective To investigate the effect of CYP3A5 on the proliferation of pancreatic cancer cells and its underlying mechanisms.Methods The protein expression of CYP3A5 in five pancreatic cancer cell lines BxPC-3,FG,MDA28,8902 and PANC1 was detected by Western blotting.The PANC1 cells with the lowest protein expression of CYP3A5 and the BxPC-3 cells with highest expression of CYP3A5 were transfected with CYP3A5 overexpression plasmid and CYP3A5 targeted-siRNA (siRNA-CYP3A5),respectively.CCK-8 and cloning formation assay were used to investigate the role of CYP3A5 overexpression and knockdown in the proliferation of pancreatic cancer cells.The changes of the protein and mRNA expression of cell cycle regulating gene cyclin E,cyclin D1 and apoptosis related gene Bcl-2 were detected by Western blotting and PCR,respectively.Results CYP3A5 protein expression in PANC1 cells increased significantly after the transfection of CYP3A5 overexpression plasmid (1.66 ± 0.14 to 1,P =0.0021),which greatly decreased in BxPC-3 cells transfected with siRNA CYP3A5 (0.18 ± 0.02 to 1,P ＜0.0001).A450 values of the CYP3A5 overexpression group and the empty plasmid group in PANC1 cells cultured for 48 and 72 h were 1.36 ±0.05 vs 1.15 ± 0.03,2.1 ± 0.09 vs 1.42 ± 0.03,respectively,which were significantly higher in CYP3A5 overexpression group than empty plasmid group,and the differences were statistically significant (P value ＜ 0.005 or 0.001).The A450 values of BxPC-3 cells in CYP3A5-siRNA transfected group and siRNA-NC transfected group were 0.62 ±0.01 vs 0.77 ± 0.03、0.83 ± 0.01 vs 1.18 ± 0.02,respectively,which in The CYP3A5-siRNA transfection group was significantly lower than that of siRNA-NC transfection group,and the difference was statistically significant (P ＜ 0.05 or ＜ 0.001).The clone formation rate of PANC1 cells in the overexpression group was (19.33 ± 0.58)％,which was significantly higher than that in the empty plasmid group (9.67±0.63) ％,and the clone formation rate in CYP3A5-siRNA group was (8.5± 0.8)％,which was significantly lower than that of group siRNA-NC (16± 0.6)％,and the differences were statistically significant (P ＜0.01).The protein expression of cyclin D1 in CYP3A5 overexpression PANC1 cells was 2.00 ± 0.11,which was obviously higher than 1.00 in empty plasmid group (P ＜0.01).The protein expression of cyclin D1 in siRNA CYP3A5 BxPC cells was 0.45 ±0.04,which was obviously lower than 1.00 in siRNA NC group,and the difference was statistically significant (P＜0.01).However,CYP3A5 overexpression or inhibition did not influence the relative expression of cyclin D1 mRNA and cyclin E,Bcl-2 pretein expression.Conclusions CYP3A5 can promote the proliferation of pancreatic cancer cells by up-regulating cyclin D1 protein expression.
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Objective To study the curative effects of jejunal exclusion surgery for STZ-induced T2DM SD rats.Methods 60 SD rats were induced to be the T2DM SD rats by intraperitoneal injection of streptozotocini.As a result,55 T2DM SD rats were successfully acquired which were randomly divided into 3 groups,20 rats in the jejunal exclusion group (A),20 rats in the sham operation group (B) and 15 rats in the control group (C).Jejunal exclusion surgery was performed in group A,jejunojejunostomy was performed in group B,and group C were fed normally.The body weight,fasting blood glucose,fasting plasma insuhn level and GLP-1 level were measured before operation and at the 1st,2rid,4th,8th and 16th week after operation.Results As compared with that before operation and that of the control group,the body weight in group A markedly declined at the 2nd,4th,8th and 16th week (352.14±9.00,342.84±8.90,336.64±10.26,330.34±9.12,P＜0.05).The fasting plasma glucose levels in group A markedly declined at the 2nd,4th,8th and 16th week (14.62±1.10,12.12±1.38,8.75± 1.06,7.55±1.00,P＜0.05).The fasting plasma insulin level in group A was maikedly increased at the 2nd,4th,8th and 16th week (14.62±3.10,16.12±3.38,17.75±4.06,17.55±3.10,P＜0.05).GLP-1 level in group A was markedly increased at the 1st,2nd,4th,8th and 16th week (11.02±0.85,14.42±1.18,16.02±1.59,17.62±1.02,18.12±0.71,P＜0.05).Conclusions The jejunal exclusion surgery is effective in controlling blood glucose,which is an ideal and lasting method.This surgery has also showed influence on body weight.
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Objective To explore the lethal action and possible mechanism of RI-1, a RAD51 inhibitor, on MSH2 deficient colorectal cancer cells.Methods The expression of MSH2 protein level was assessed by Western blot, and the sensibility of human colorectal cancer cells to RI-1 (10, 20, 30, 40 and 50 μmol/L)was measured by MTT method.Lentivirus vectors MSH2-shRNA and Neg-shRNA (negative control) were constructed and transfected into HT29 cell.Apoptosis and DNA damage of cells treated with RI-1(40 μmol/L)were detected by flow cytometry and Single cell gel electrophoresis respectively.In addition, the formation of γ-H2AX foci was analyzed by immunofluorescence.Results Compared with control, MSH2-deficient HCT8 cells had obviously apoptosis(P<0.01);in HCT8 and HT29 Shmsh2 cells, tail DNA%, tail length, tail moment and olive tail moment were markedly increased(P<0.05),and the number of γ-H2AX focus were increased(P<0.01).Conclusions RAD51 inhibitor RI-1 selectively kills MSH2 deficient colorectal cancer cells by increasing DNA damage.
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Interactions between noroviruses (NoVs) and the receptors of histo-blood group antigens (HB-GAs) affect the infectivity and host susceptibility of NoVs. We elucidated the binding profile of a GII. 12 NoV to HBGAs. First, we synthesized the P domain sequence of the GII. 12 NoV strain Pune (GenBank accession number EU921353). Protein of the P domain was expressed in a prokaryotic system. Formation of the P particle was monitored by gel-filtration chromatography. Antiserum was prepared by immunization of mice with GII. 12 P particles. The binding profile of the GII. 12 NoV Pune strain was determined by binding of the P particle with a panel of saliva samples with various known HBGAs phenotypes. The GII. 12 NoV was bound strongly to saliva samples of subjects with B and AB types and weakly to A, O secretor, and non-secretor saliva samples, suggesting higher affinity with B antigen by GII. 12 NoV. These results were consistent with those determined by a previous crystallography study of GII. 12 NoV. These data suggested that individuals with B and AB blood types may be more susceptible to infection by GII. 12 NoV compared with those with other blood types. Our findings may provide a basis for the prevention and control of an epidemic of GII. 12 NoV.
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Animals , Female , Humans , Mice , Blood Group Antigens , Metabolism , Caliciviridae Infections , Metabolism , Virology , Gastroenteritis , Metabolism , Virology , Genotype , Mice, Inbred BALB C , Norovirus , Genetics , Metabolism , Protein Binding , Receptors, Virus , Metabolism , Viral Proteins , Genetics , MetabolismABSTRACT
Enterovirus 71 (EV71) is a major agent of hand, foot and mouth disease that can cause a severe burden of disease to children. To identify an effective method for the control and prevention of EV71, we studied the effect of exposure to heat and ultraviolet (UV) light upon EV71 inactivation. We found that exposure to 50 degrees C could not inactivate the infectivity of EV71. However, exposure to 60 degrees C and 70 degrees C could inactivate EV71 effectively. EV71 could be inactivated after exposure to UV light at a distance between the sample and a lamp of 30 cm for 30 min or 60 min because viral genomic RNA was destroyed. However, fetal bovine serum (FBS) could attenuate the inactivation proffered by heat and UV light. Attenuation effects of FBS were correlated positively with FBS concentration. Hence, EV71 can be inactivated by exposure to heat and UV light, and our results could provide guidance on prevention of the spread of EV71.
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Humans , Disinfection , Methods , Enterovirus A, Human , Genetics , Physiology , Radiation Effects , Enterovirus Infections , Virology , Hot Temperature , Ultraviolet Rays , Virus Inactivation , Radiation EffectsABSTRACT
Chaihu Injection (CI), which is widely used in treatment of febrile diseases, is an aqueous solution of Chaihu (Radix Bupleuri Chinensis) or Nanchaihu (Radix Bupleuri Scorzonerifolii) prepared by steam distillation.
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Aiming at the nonlinear system identification problem, a parallel recursive affine projection (AP) adaptive algorithm for the nonlinear system based on Volterra series is presented in this paper. The algorithm identifies in parallel the Volterra kernel of each order, recursively estimate the inverse of the autocorrelation matrix for the Volterra input of each order, and remarkably improve the convergence speed of the identification process compared with the NLMS and conventional AP adaptive algorithm based on Volterra series. Simulation results indicate that the proposed method in this paper is efficient.
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BACKGROUND: Radix astragali has the effect of protecting cells from damage in ischemic reperfusion, whether pre-treatment with radix astragali can protect myocardial eells from apoptosis in ischemic reperfusion ? OBJECTIVE: To investigate the effect of pre-treatment with radix astragali on apoptosis and its relative genes in rats with ischemic myocardial reperfusion DESIGN: A randomized and controlled trial taking Wistar rats as experimental subjects.SETTING: The Basic Medical Department of Chengde Medical College and the Geriatric Department of the Affiliated Hospital.MATERIALS: The experiment was completed in the Imunnohistochemical Laboratory of Basic Medical Institute in Chengde Medical College from February to December in 2004. A total of 30 healthy male Wistar rats were selected, and at random classified as groups of radix astragali pre-treated (radix astragali), ischemic reperfusion and psuedo-operated (control), 10 rats for each group.METHODS: Radix astragali injection was given peritonealy for rats in radix astragali pre-treated group before operation, and the equivalent normai saline was given for those in ischemic reperfusion and psuedo-operated groups. One week later, the model of ischemic reperfusion was set up. After operation the myocardia in marginal zone of ischemic reperfusion were sampled, and the myocardia of the corresponding zone were taken for control group. The method of terminal (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) was used for assay of myocardial apoptosis rate, and the ABC immunohistochemical method was used for assay of myocardial bcl-2 (inhibiting apoptosis gene) and bax (promoting apoptosis gene).MAIN OUTCOME MEASURES: Apoptosis rates, and expression of bcl2 and bax genes of myocardia RESULTS: ① Apoptosis rate of myocardial cells: The rate in radix astragali group was decreased compared with that in ischemic reperfusion group [ (14.06 ±9.97) ％, (19.34±12.30) ％, t = 1.863, P ＜ 0.05].② Expression of bcl-2: There was no significant difference between radix astragali and ischemic reperfusion groups[(9.14±4.46) ％, (8.99±4.54) ％, P ＜ 0.05].③ Expression of bax: The expression in radix astragali group was decreased compared with that in ischemic reperfusion group [(12.65 ±7.23)％,(18.12±7.92) ％, t = 2.096, P ＜ 0.05]CONCLUSION: Pre-treatment with radix astragali can down-regulate the expression of promoting apoptosis gene so as to reduce the rate of myocardial cell apoptosis, hence it can protect the myocardial cells in ischemic reperfusion.
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Objective The microvasculature of the anterior lobe, posterior lobe and cerebellar vermis was observed under the optical microscope in 6 rats. Methods The tannic acid-ferric chloride method (TAFM) was used to stain the vessels in cerebellum. Results The arteries within the cortex and medulla of the cerebellum originated from pia mater arteries and central branches of the cerebellar arteries, the branches of pia mater arteries mostly penetrated into the cerebellar cortex vertically and formed capillary net in the molecular layer, purkinje layer and granular layer, the arteriole in the medulla of cerebellar posterior lobe branched and formed clawed-like figurations. These vessels beneath lobule cortex connected mutually by vessel anatomosis across the medulla. Conclusion The arteriole in the cerebellum can be manifested distinctly in a strong three-dimension by TAFM and the vessel anatomosis in deep layer of cerebellum can be easily observed.