Objective:To construct a small interfering RNA(siRNA)expression vector(psiRNA-VEGFR-3)targeting vascular endothelial growth factor receptor 3(VEGFR-3)and to investigate the effects of VEGFR-3 siRNA on the adherence and invasion of human colon cancer cells.Methods:A siRNA expression vector(psiRNA-VEGFR-3)targeting VEGFR-3 were constructed and was used to transfect LoVo cells via lipofectamine 2000.The mRNA and protein expression of VEGFR-3 were examined after transfection by reverse transcriptase polymerase chain reaction(RT-PCR)and Western blotting,respectively.The tumor adhesion ability was detected by cell-matrix adhesion experiment and the invasion ability of tumor cells was evaluated by millicell chamber model.Results:The VEGFR-3 siRNA expression vector was successfully constructed.The expression of VEGFR-3 mRNA and protein was inhibited after psiRNA-VEGFR-3 transfection.Seventy-two hours after psiRNA-VEGFR-3 transfection,Western blotting assay showed that the expression of VEGFR-3 protein was decreased from(1.26?0.19)to(0.39 s0.12)(P

Objective To study the relative factors of hypotension after recanalization of infarction related artery(IRA) during emergency PCI for patients with acute myocardial infarction.Methods We retrospectively analyzed the clinical information and the coronary angiograph of 245 consecutive patients with primary acute myocardial infarction undergoing emergency PCI at the first affiliated hospital of China Medical University from January 2003 to September 2006.Twenty seven patients with blood pressure elevated higher than 140/90 mm Hg after reperfusion were excluded.The remaining 218 patients were catagorized into the hypotension group(BP≤90/60 mm Hg,n=102) and the normal blood pressure group(n=116) according to the blood pressure level after reperfusion during emergency PCI.The relative factors of hypotension were analyzed through logistic regression test.Results Compared with the normal blood pressure group,patients in the hypotension group had higher incidences of old age(age ≥60 yrs),heart failure(killip grades≥grade 2),inferior wall or right ventricular infarction,right coronary artery lesions,proximal or total occlusion lesions,multivessal disease and TIMI blood flow≤grade 1 in the IRA after recandization.Mortality rate was also higher in the hypotension group(P1).Conclusion Old age(age ≥60),inferior wall infarction,killip grades≥2,right coronary artery lesions,proximal or total occlusion lesions are the risk factors of hypotension after recanalization of IRA during emergency PCI.The presence of hypotension is correlated with the increase of in hospital mortality.

Objective To study the practicality and repeatability of B-mode ultrasonography on diagnosis and score of carotid atherosclerosis. Methods Ninety-nine B-mode ultrasonography pictures of carotid atherosclerosis were obtained from field investigation. According to the established standard(normal scored as 0, other scored as 1 - 7 by the severity), the 99 pictures were diagnosed and scored once together by three trained inexperienced members of the research team. Ten days later, these pictures were diagnosed and scored respectively, by the three members.The diagnostic results were analyzed with statistical methods to test the feasibility and repeatability of this set of diagnostic and scoring system. Results The joint diagnostic results by the three group members were regard as standard with scores 0 in 27 cases, 1 in 27 cases, 2 in 10 cases, 3 in 4 cases, 4 in 13 cases, 5 in 8 cases, 6 in 6 cases, and 7 in 4 cases. Independent diagnostic results by member A was 0 in 29 cases, 1 in 24 cases, 2 in 11 cases, 3 in 2 cases, 4 in 12 cases, 5 in 11 cases, 6 in 6 cases, and 7 in 4 cases. Independent diagnostic results by member B was 0 in 29 cases, 1 in 22 cases, 2 in 14 cases, 3 in 4 cases, 4 in 13 cases, 5 in 5 cases, 6 in 8 cases, and 7 in 4 cases. Independent diagnostic results by member C was 0 in 28 cases, 1 in 25 cases, 2 in 8 cases, 3 in 5 cases, 4 in 14 cases, 5 in 10 cases, 6 in 4 cases, and 7 in 5 cases. Comparison of the 4 diagnostic scores, the difference was not statistically significant(F = 0.019, P ＞ 0.05). Joint diagnostic results of the three members were compared with the standard and correlation coefficient were 0.977,0.987,0.932, respectively(all P ＜ 0.01 ). The correlation coefficient of diagnostic results of each member were 0.969,0.935,0.928, respectively (all P ＜0.01 ). Diagnostic results of each member were consistent with the standard and the compliance rate were 88.9% (88/99) ,90.9%(90/99) and 86.9%(86/99), respectively. Conclusions B-mode ultrasonography is a non-injure method for diagnosis of carotid atherosclerosis in epidemiology investigation. The method is easy to grasp and has a good repeatability.

Objective To investigate the possible protective effect of neurotropin on the corresponding spinal cord motoneuron after sciatic nerve injury early in rats.Methods 108 male Sprague Dawley rats were divided randomly into Groups A,B and C with 36 animals in each group,and all animals were cut both side of sciatic nerve,instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve.Group A was control group.Group B was treated with neurotropin and Group C was treated with normal saline after operation.The rats were sacrificed on 1st d,4th d,9th d,14th d,21st d and 28th d after operation.The corresponding spinal cord segments(L4～L6)were gotten to make Bcl-2 and Bax immunohistochemical staining and TUNEL staining.Results The values of Bcl-2/Bax on 9th d,14th d and 21st d after operation of Group B were different from that of Group A and Group C.The number of positive cells stained by TUNEL staining on 9th d was significantly different among Group A,Group B and Group C(v<0.05).The expression of positive TUNEL staining cells reached a peak on 14th d after operation,the number of Group B was less than that of Group A and Group C.Conclusion After instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve,neurotropin can protect the spinal cord motoneurons to avoid apoptosis excessively in some extent.

Objective To study relationships between myocardial injury and the levels of serum complement C3, C4 and C5b-9 in patients with acute coronary syndrome (ACS). Methods A retrospectively analysis was conducted. 170 ACS patients [including 110 cases of ST-segment elevation myocardial infarction (STEMI) and 60 cases of non-ST-segment elevation acute coronary syndrome (NSTE-ACS)] with ischemic chest pain or chest discomfort onset within the prior 12 hours admitted to the cardiology department of Tianjin Union Medicine Center from January 2014 to July 2016 were enrolled. Thirty-six healthy cases were enrolled as control during the same time. The levels of serum complement C3, C4 and C5b-9 on 1, 3 and 7 days after admission and myocardial function indicators were analyzed. Major adverse cardiovascular events (MACE) and readmission rate were analyzed after 1 year follow-up. The correlation between serum complement levels and myocardial function indicators was analyzed by Pearson correlation analysis. Results ① The levels of serum C3, C4 and C5b-9 on the first day in NSTE-ACS group and STEMI group were significantly higher than control group [C3 (g/L): 1.04±0.33, 1.26±0.35 vs. 0.39±0.21, C4 (g/L): 0.31±0.14, 0.33±0.10 vs. 0.19±0.07, C5b-9 (g/L): 575.46±197.26, 659.26±160.77 vs. 501.40±141.51, all P < 0.05]. There were no changes of serum C3, C4 in NSTE-ACS group, but C5b-9 decreased after a peak (g/L: 700.63±218.42) at 3 days. Serum complements in STEMI group reached peak on the third day [C3 (g/L): 1.37±0.33, C4 (g/L): 0.42±0.12, C5b-9 (g/L): 754.72±136.22]. The levels of serum C4 and C5b-9 in STEMI group were higher than NSTE-ACS group on the third and seventh day. ② The levels of troponin T (TnT), creatine kinase-MB (CK-MB), solution intercellular adhesion molecule-1 (sICAM-1), global registry of acute coronary events (GRACE) scores and percutaneous coronary intervention (PCI) numbers in STEMI group were significantly higher than those in the NSTE-ACS group, which were as opposite as left ventricular ejection fraction (LVEF). However, there were no significant differences in levels of serum N-terminal pro-brain nitric peptide (NT-proBNP), Fibrinogen (Fib), readmission rate and incidence of MACE between STEMI and NSTE-ACS groups. ③ According to GRACE, patients with ACS were divided into low risk group (≤ 108 scores, 26 cases), intermediate risk group (109-140 scores, 61 cases) and highest group (> 140 scores, 83 cases). TnT and sICAM-1 in intermediate risk group were significantly increased as compared with low risk group. Levels of TnT, sICAM-1, C3, C4 and C5b-9 in the highest group were significantly higher than the low and intermediate risk groups, however the lowest LVEF was found in the highest group. ④ It was shown by Pearson correlation analyses that levels of serum C3, C4, C5b-9 were positively correlated with TnT (r value was 0.481, 0.367, 0.292, respectively, all P <0.01), sICAM-1 (r value was 0.298, 0.249, 0.365, respectively, all P < 0.01), but negatively correlated with LVEF (r value was -0.384, -0.260, -0.200, respectively, all P < 0.01). In addition sICAM-1 positively correlated with TnT (r = 0.536, P = 0.000), but negatively correlated with LVEF (r = -0.341, P = 0.001). Conclusions Serum complements activation was found in the acute phase of ACS patients. Serum complement C3, C4 and C5b-9 are involved in the process of myocardial injury, and may reflect severity of myocardial injury and cardiac dysfunction.

Animals ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-18 ; biosynthesis ; genetics ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Analysis ; Time Factors ; Transfection ; Transplantation, Heterologous

Objective To observe the effects of dexmedetomidine hydrochloride on the cisatracurium-induced neuromuscular blockade in geriatric, young and middle-aged patients. Methods Forty elderly patients and forty young and middle-aged patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 4 groups ( n=20 each):elderly dexmedetomidine hydrochloride group (group DE),elderly control group (group CE),young and middle-aged dexmedetomidine group (group DY) and young and middle-aged control group (group CY).In groups DE and DY,a loading dose of 0.5 μg?kg-1 dexmedetomidine was intravenously infused over 10 min before induction of anesthesia,respectively,followed by infusion at a rate of 0.4μg?kg-1?h-1 until the end of surgery.Equal volume of 0.9% sodium chloride was given in groups CE and CY.Anesthesia was induced with midazolam,fentanyl,propofol and cisatracurium 0.15 mg?kg-1.Four groups were maintained with infusion of propofol and remifentanil. Neuromuscular blockade was maintained with continuous cisatracurium infusion in the four groups and was monitored with TOF-Watch SX acceleromyography at the adductor pollicis.The onset time,TOF no reaction period,duration of action,the amount of cisatracurium consumption,and the spontaneous recovery index ( T1 25% to 75%) were recorded. Results The four groups were comparable in the demographic data.Intubation conditions,the onset time and recovery index were not significantly different among the four groups. The duration of blockade action in groups DE and DY was ( 61. 1 ± 8.9) min and (53.6±9.3) min,which was significantly longer than that in group CE [(49.9±5.8) min] and group CY [(44.8± 6.4) min] (P<0.01).The duration of blockade action was significantly longer in groups DE and CE than in groups DY and CY (P<0.05).The amount of cisatracurium consumption in groups CE and CY was significantly higher than that in groups DE and DY (P<0.05). Conclusion The neuromuscular blockade is longer and the requirement of cisatracurium is less in elderly patients than in young and middle-aged patients.Continuous infusion of dexmedetomidine hydrochloride can't accelerate the onset time of cisatracurium.But duration of action is prolonged and the amount of cisatracurium consumption is lower in patients with infusion of dexmedetomidine hydrochloride.

Objective To evaluate the effect of noninvasive ventilation (NIV) on hypercapnic encephalopathy syndrome (HES) induced by acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods An extensive search of related literature from the PubMed, EMBASE, Cochrane library, CNKI and Wanfang databases up to January 2015 was performed. Randomized controlled trials (RCTs) and case control studies regarding comparison of the effect of NIV and conventional mechanical ventilation (CMV) on the HES were collected. Critical appraisal skills program (CASP) was adopted to assess the quality of the studies. Data including mortality, trachea intubation rate, duration of mechanical ventilation and complication rate were collected, and Meta-analysis was performed by RevMan 5.3. Results Finally, 6 studies were included with 225 subjects, among whom 112 were in NIV group and 113 in CMV group, and the average Kelly-Matthay score was 3. Compared with CMV group, the mortality [20.5% (23/112) vs. 32.7% (37/113), risk ratio (RR) = 0.63, 95% confidential interval (95%CI) = 0.40-0.98, P = 0.04], intubation rate [35.7% (40/112) vs. 100.0% (113/113), RR = 0.38, 95%CI = 0.26-0.55, P < 0.000 01], incidence of ventilation related complications [26.2% (21/80) vs. 50.6% (42/83), RR = 0.52, 95%CI = 0.34-0.79, P = 0.002] in NIV group were significantly decreased, and the duration of mechanical ventilation was significantly shortened [days: 7.1 vs. 16.2, standard mean difference (SMD) = -0.93, 95%CI = -1.39 to -0.46, P < 0.000 1]. Conclusion NIV could significantly lower the mortality rate, intubation rate, and complications in the treatment of HES induced by AECOPD under close monitoring.

Objective To investigate the mechanism of drug resistance mediated by micF gene and outer membrane porin F ( OmpF) in Shigella strains. Methods Shigella strains were isolated from stool samples of patients who presented to the Second Hospital of Tianjin Medical University with acute diar-rhea in 2015. Antibiotic susceptibility test was performed to screen out the multidrug-resistant and non-multi-drug-resistant strains. The ompF gene was amplified by PCR. The micF and ompF genes at transcriptional levels in the two groups of strains were detected by quantitative real-time RT-PCR. Intracellular concentra-tions of ciprofloxacin in the two groups of Shigella strains were measured by automatic microplate reader. Re-sults According to the result of antibiotic susceptibility test, 13 strains that were resistant to 3 or more than 3 antibiotics were classified into the multidrug-resistant group, while the other 8 strains that were sensitive to all antibiotics used in this study or only resistant to 1 or 2 antibiotic were classified into the non-multidrug-re-sistant group. All of the 21 Shigella strains carried the ompF gene. Compared with the non-multidrug-resist-ant strains, the multidrug-resistant strains showed higher expression of micF gene, but lower expression of ompF gene. The differences in micF and ompF genes between the two groups were statistically significant. The result of correlation analysis suggested that there was a negative correlation between micF and ompF genes (r=-0. 244). The intracellular concentrations of ciprofloxacin in multidrug-resistant strains were low-er than those in the non-multidrug-resistant strains (P<0. 001). Conclusion The decreased expression of OmpF was one of the possible mechanisms of multidrug-resistance in Shigella strains. The micF gene was negatively related to the expression of OmpF. Moreover, the decreased intracellular concentrations of cipro-floxacin in multidrug-resistant strains might be related to the decreased expression of OmpF.

Objective To detect the osteopontin(OPN)expression in renal tissue of rats with fluorosis and low calcium diet,and study the role of OPN in renal injury of fluorosis.Methods Forty-eight aged 1 month Wistar rats,80-120 g,were randomly divided into 4 groups by 2×2 factorial design(the number of female and male in each group was equal):the control group,high-flluoride group,low-calcium group and low-calcium with high-fluoride group.All rats of the fluorosis groups were fed with feed containing corn exposed to coal-burning from endemic fluorosis areas with high fluoride(100 mg/kg,corn),the other two groups were fed with feed containing coru from nonendemic fluorosis areas(fluoride 5 mg/kg,corn).After 16 weeks,the rats were killed.The change of teeth was examined,and the incidence rate of dental fluorosis was calculated.The expressions of both protein and mRNA of OPN in rat renal tissue were determined by RT-PCR and immunohistochemistry after four-month experimentation.Results The growth of teeth was very well in the control group and the low-calcium group.The two high-fluoride groups showed evident dental fluorosis(100%).The results of immunohistochemistry showed that the OPN protein was localized in renal tubule cytoplasm.The OPN-positive cells from renal tissue were lightly and scatteredly stained in control and low-calcium groups.The OPN-positive cells had deeper color in high-fluoride group and low-calcium with high-fluoride group,widely distributed in the renal tubular epithelial cells.The protein expression of OPN in the two groups exposed to fluoride(168.64±13.21,169.26±8.92)was significantly higher than those of the corresponding control group(145.78±10.26,all P<0.01)and low-calcium group(149.60±16.84,all P<0.01).The mRNA expression of OPN in the two groups exposed to fluoride(1.89±0.37,1.94±0.22)was significantly higher than those of the corresponding control group(1.32±0.26,all P<0.05)and low-calcium group(1.30±0.186,P<0.05),respectively.High fluoride influenced the expression of protein and mRNA of OPN(F=13.821,4.24,all P<0.05).Low calcium did not affect the expression of protein and mRNA of OPN(F=2.164,0.58,all P>0.05).However,high fluoride and low calcium had a cross interaction on the expression of protein and mRNA of OPN(F=6.257,432,all P<0.05).Conclusions Over-dose fluoride enhances the expression of OPN.The higher expression observed in the cases exposed to high fluoride concentration is associated with serious renal injury.OPN may he a potential marker for renal injury in fluorosis.Moreover,over-dose fluoride and low calcium make the renal injury worse,indicating low calcium plays an important part in renal injury by fluoride.