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Objective:To investigate the effects of the non-steroidal anti-inflammatory drugs Parecoxib and Flurbiprofen administered at different time points on mesenteric traction syndrome(MTS).Methods:This was a prospective, randomized, controlled clinical trial.One hundred elderly patients scheduled for open radical gastrectomy under general anesthesia were randomly allocated to four groups: the control group, the P-Pre-MT group, the F-Pre-MT group, and the F-Post-MT group(n=25, each group). Parecoxib 40 mg and Flurbiprofen 50 mg were intravenously administered 30 min and 5 min before skin incision in the P-Pre-MTS group and the F-Pre-MTS group, respectively.Flurbiprofen 50 mg was infused at the moment of MTS in the F-Post-MTS group while the control group was intravenously injected with saline.Anesthesia induction and maintenance were performed with plasma target-controlled infusion of Propofol and Remifentanil.After the incision of the peritoneum.The incidence of MTS, the duration of hypotension, and the use of norepinephrine during MTS were recorded.Systolic blood pressure(SBP), heart rate(HR), and effect-site concentration of Remifentanil were monitored at MT(T 0), 10 min(T 10), 20 min(T 20), 30 min(T 30), 45 min(T 45), and 60 min(T 60)after MT in patients with MTS. Results:MTS was observed in 19 of 22 patients(86%), 19 of 23 patients(83%), 0 of 24 patients(0%)and 20 of 23 patients(87%)in the control, P-Pre-MT, F-Pre-MT and F-Post-MT groups, respectively.The incidence of MTS in the F-Pre-MT group was lower than that in the control group( χ2=35.313, P=0.000). The duration of hypotension and the use of norepinephrine in patients with MTS were less in the F-Post-MT group than in the control group( P=0.007 and 0.015). SBP and HR at different time points after MT had significant differences in patients with MTS in the control group( F=47.425 and 26.318, P=0.000 and 0.000), but did not differ in the F-Pre-MT group( F=2.140 and 1.013, P=0.066 and 0.413). SBP and the effect-site concentration of Remifentanil were lower and HR was higher in the control group than in the F-Pre-MT group at T 10and T 20after MT( P=0.000), and SBP was higher and HR was lower in the F-Post-MT group than in the control group C at T 20after MT( P=0.002 and 0.002). Conclusions:Flurbiprofen not only can prevent the occurrence of MTS, maintain blood pressure stability and heart rate after MT, but also can reduce the duration of hypotension and the amplitude of heart rate increase when MTS occurs in elderly patients undergoing open radical gastrectomy.Parecoxib has no effect on MTS.
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Objective To evaluate the role of calcineurin ( CaN)/nuclear factor of activated T cell cytoplasmic 4 protein ( NFATc4) signaling pathway in inflammatory responses in lung tissues of rats with ventilator-induced lung injury ( VILI) . Methods Twenty-four clean-grade healthy male Wistar rats, aged 5-8 weeks, weighing 220-250 g, were divided into 3 groups ( n=8 each) using a random number table method: control C (group C), VILI group and cyclosporine A plus VILI group (group CsA+VILI). The animals were anesthetized with pentobarbital and tracheostomized. The rats were mechanically ventilated for 6 h with the tidal volume set at 40 ml/kg and respiratory rate at 40 breaths/min to establish the model of VI-LI. The rats kept spontaneous breathing in group C. CaN specific inhibitor cyclosporine A 10 mg/kg was in-traperitoneally injected at 1 h before ventilation in group CsA+VILI. Rats were sacrificed immediately after ventilation, lung tissues were obtained and stained with hematoxylin and eosin to evaluate lung injury, broncho-alveolar lavage fluid was collected for determination of tumor necrosis factor-alpha ( TNF-α) , inter-leukin-1beta ( IL-1β) and IL-6 concentrations by enzyme-linked immunosorbent assay, and the lungs were removed for determination of the wet to dry weight ratio ( W/D ratio) , expression of intercellular adhesion molecule-1 ( ICAM-1) and vascular cell adhesion molecule-1 ( VCAM-1) ( by real-time polymerase chain reaction) , and expression of calcineurin and NFATc4 in lung tissues ( using Western blot ) . Results Compared with group C, the W/D ratio, lung injury scores and concentrations of IL-1β, IL-6 and TNF-αin BALF were significantly increased, and the expression of CaN, NFATc4, ICAM-1 mRNA and VCAM-1 mRNA was up-regulated in group VILI ( P＜0. 05) . Compared with group VILI, the W/D ratio, lung injury scores and concentrations of IL-1β, IL-6 and TNF-αin BALF were significantly decreased, and the expres-sion of CaN, NFATc4, ICAM-1 mRNA and VCAM-1 mRNA was down-regulated in group CsA+VILI ( P＜0. 05) . Conclusion CaN/NFATc4 signaling pathway mediates inflammatory responses in lung tissues of rats with VILI.
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Humanistic care centered on the interests and needs of patients, is an important means to maintain the doctor-patient relationship and to build a harmonious society. However, there are still some problems in the major public hospitals, such as inadequate communication and coordination, complicated and time-consuming medical procedures, and so on. Based on this, this paper discussed the problems existing in public hospitals in Anhui Province, and put forward some measures, such as strengthening doctor-patient communication, caring for doctor-patient personnel, and constructing hospital culture, so as to perfect the humanistic care system.
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Objective To determine the optimal pressure for facemask ventilation during induction of general anesthesia by real-time ultrasonographic measurement of antral cross-sectional area (CSA) in adult patients.Methods Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 18-60 yr,with body mass index of 20-25 kg/m2,scheduled for elective operation under general anesthesia,were divided into 5 groups (n=12 each) using a random number table:P10 group,P13 group,P16 group,P19 group and P22 group.After induction of anesthesia,an oropharyngeal airway was inserted,and the patients were ventilated for a 2-min period in a pressure-controlled mode using the two-handed mask ventilation technique.The pressure for facemask ventilation was 10,13,16,19 and 22 cmH2O in P10,P13,P16,P19 and P22 groups,respectively.The antral CSA was measured using real-time ultrasonography before and after facemask ventilation.Respiratory parameters were recorded.Results Compared with group P1O,the number of patients in whom CSA＜340 mm2 after facemask ventilation was significantly decreased in P16,P19 and P22 groups,and the number of patients in whom the tidal volume ≥ 6 ml/kg was increased in P13,P16,P19 and P22 groups (P＜ 0.01).The number of patients in whom optimnal pressure for facemask ventilation was achieved was 2,10,6,4 and 1 in P10,P13,P16,P19 and P22 groups,respectively,with the most cases in group P13 (P ＜ 0.01).Conclusion The optimal pressure is 13 emH2O for facemask ventilation during induction of general anesthesia when determined by realtime ultrasonographic measurement of antral CSA,and it can ensure adequate oxygen supply and reduce gastric insufflation in adult patients.
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Objective To explore the effect of dexmedetomidine on blood coagulation following radical gastrectomy.Methods ASA Ⅰ or Ⅱ patients aged 51-70 years weighing 53-75 kg scheduled for radical gastrectomy were randomly allocated to two groups:dexmedetomidine group (group D)and control group (group C).Dexmedetomidine 0.5 μg/kg was intravenously infused over 10 minutes before anesthesia induction,followed by a rate of 0.5 μg·kg-1 ·h-1 until peritoneal closure in group D and volume-matched normal saline was administrated in group C.Radical gastrectomy was performed under total intravenous anesthesia with propofol and remffentanil.A series of warming measures were implemented and artificial colloid and heparin flushing fluid were not used.Postoperative patient-controlled intravenous analgesia was performed to maintain visual analogue scale≤3.The blood samples were collected for TEG and standard coagulation monitoring before dexmedetomidine and saline administration and 3 h after surgery.Results The temperature and hematocrit in the postoperative period were significantly less than the preoperative period in two groups (P＜0.01).In both groups,the activity of plasma antithrombin Ⅲ was significantly decreased and the concentration of plasma FDP was significantly increased in the postoperative period when compared with the preoperative period (P ＜0.01).In group D,the R time was significantly shortened and MA value was significantly increased in the postoperative period when compared with the preoperative period (P＜0.05) and there were no significant differences in the K time and α angle between the preoperative and postoperative period.In group C,the R and K time were significantly shortened and the value for MA and α angle were significantly increased in the postoperative period compared with the preoperative period (P＜0.01).The platelet counts,PT,APTT,and plasma fibrinogen concentration were comparable between the preoperative and postoperative period in both groups.The requirements of propofol and remifentanil in group D were significantly less than group C (P＜0.05).In the preoperative period,the plasma antithrombin Ⅲ activity,FDP concentration,and the values for all TEG variables were similar in both groups.In the postoperative period,the value for MA and the concentration of plasma FDP in group D were less than that in group C and the value for R and the activity of plasma AT Ⅲ in group D were more than group C (P＜0.05 or P＜0.01) and there were no significant differences in the K time and α angle in both groups.There were no significant differences in the temperature,hematocrit,platelet counts,PT,APTT,and plasma fibrinogen concentration in the preoperative and postoperative periods between the two study groups.Conclusion Adjunctive dexmedetomidine in general anesthesia could inhibit the decrease of R time and raise of the value for MA,inhibit the decrease of plasma an tithrombin Ⅲ activity and raise of FDP concentration,which indicated that dexmedetomidine can improve blood coagulation state after radical gastrectomy.
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Objective To develop a novel method for genotyping rs 738409 ( C >G ) single nucleotide polymorphism ( SNP ) , and explore the association between the rs 738409 genotypes and non-alcoholic fatty liver disease ( NAFLD ) .Methods Method establishment and analysis of genetic susceptibility.The principle of amplification refractory mutation system ( ARMS) and combined a TaqMan fluorogenic probe as signal report were used , by monitoring the difference of cycle threshold (ΔCt=C allele-special primer Ct values minus G allele-special primer Ct values ) between the two PCR reactions in a real-time PCR, the method for rs738409 genotyping was established ( ARMS-TaqMan) .618 subjects ( men:401;women: 217 ) , in an annual health check-up program from January 2011 to December 2014 in Aerospace Center Hospital , were performed for rs738409 genotyping by the ARMS-TaqMan assay.Fatty liver was diagnosed based on abdominal ultrasonography .The chi-square test and multiple logistic analyses were used to analyze the relationship of rs 738409 genotypes and NAFLD.Results The ΔCt by ARMS-TaqMan for rs738409 genotyping were -13.1 ±1.4 of CC alleles ( 243 cases ) , 0.01 ±0.45 of CG alleles ( 282 cases), and 12.7 ±1.9 of GG alleles (93 cases), respectively.The GG alleles frequency of rs738409 were significantly higher in patients with NAFLD compared with subjects without NAFLD (21.5%vs 12.3%,χ2=8.677, P =0.003).In comparison to subjects with CC alleles, the OR (95% confidence interval) adjusted for age, gender, total cholesterol, triglyceride, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol, fasting blood glucose and body mass index was 1.35 (0.91-2.00) in subjects with CG alleles and was 2.21 ( 1.32 -3.71 ) in subjects with GG alleles ( P =0.013 ) .Variant rs738409 genotypes were associated with significant increased trend in alanine aminotransferase ( ALT) level from CC alleles, CG alleles to GG alleles (F=8.980, P<0.001), and in aspartate aminotansferase (AST) between GG alleles and CC alleles (F=6.491, P<0.001).Conclusions The novel ARMS-TaqMan assay had the features of accuracy , one step and high-throughput for rs738409 genotyping.The G allele of rs738409 was a risk factor of NAFLD susceptibility and associated with higher level serum ALT and AST .
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Objective To investigate the relationship between T?cell death?associated gene 8 ( TD?AG8) and endogenous neuron?protective mechanism against oxygen?glucose deprivation and restoration ( OGD∕R)?induced apoptosis in rat neurons. Methods The primary cortical neurons obtained from fetal rats were seeded in 6?well plates at a density of 1×105 cells∕ml and divided into 5 groups using a random number table: control group ( group C, n=24 ) , group OGD∕R ( n=48 ) , TDAG8 agonist BTB09089 group (group BTB, n=24), TDAG8?siRNA group ( group siRNA, n=24), and blank vehicle group ( group V, n=24) . The medium was replaced with glucose?and serum?free Locke′s buffer, and the neu?rons were exposed to 95% N2?5% CO2 in an air?tight incubator at 37℃ for 60 min followed by routine cul?ture to establish the model of OGD∕R. In BTB, siRNA and V groups, 20 μmol∕L TDAG8 agonist BTB09089, 200 pmol∕L TDAG8?siRNA, and 6 μl∕200 μl transfection reagent were added, respectively, at 24 h before oxygen?glucose restoration. At 6 h of oxygen?glucose restoration, the neuronal viability and a?mount of lactic dehydrogenase ( LDH) released were measured, and the expression of TDAG8 and caspase?3 mRNA in neurons was detected by fluorescent quantitative real?time polymerase chain reaction. In group OGD∕R, the expression of TDAG8 and caspase?3 was measured by Western blot at 0, 3, 6, 12 and 24 h of oxygen?glucose restoration. In C, OGD∕R, BTB, siRNA and V groups, the expression of TDAG8, caspase?3 and p?Akt was detected at 6 h of oxygen?glucose restoration. Results In group OGD∕R, the ex?pression of TDAG8 was gradually up?regulated after oxygen?glucose restoration, and the expression of caspase?3 peaked at 6 h of oxygen?glucose restoration. Compared with group C, the neuronal viability was significantly decreased, the amount of LDH released was significantly increased, and the expression of TD?AG8 and caspase?3 protein and mRNA and p?Akt was significantly up?regulated in OGD∕R, V and siRNA groups ( P0?05) . Conclusion TDAG8 is partially involved in the endogenous neuron?protective mechanism against OGD∕R?induced apoptosis in rat neurons, which may be related to activation of Akt signaling pathway.
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Objective To evaluate the application effect of the problem based learning in clinical teaching of orthopedics. Methods We chose sixty five-year program students of Peking univ-ersity health science center as research object. All students were randomly divided into experimental and control groups, with thirty students in each group. Experimental group was given PBL teaching combined with lecture based learning (section-based learning, LBL), while the control group only received the LBL teaching. Two groups were given 8 hours of teaching experiments. After the end of the study, the teaching effect of the two groups was evaluated by the theory course and clinical skills. The questionnaire was distributed to the 2 groups. The scores of both experimental group students and control group students in theory courses and clinical skills were analyzed using SPSS 19.0 software using t-test and x2 test. Result The score of the experimental group was (53.7 ±3.2) in the knowl-edge-based grades exam, while the control group was (52.3±2.2), showing no obvious difference when compared to the control group's grades (P>0.05). The score of the experimental group was (24.0±1.5) in the hands-on technique grades exam, while the control group was (22.3±1.6). The difference in grades showed statistical significance (P<0.05). Feedback survey results showed that the experimental group's teaching satisfaction degree was also significantly higher than the control group, and the dif-ference was statistically significant (P<0.05). Conclusions The application of PBL in clinical teach-ing in orthopedic clinical teaching can improve students' learning interest and self-study ability, and helps to develop students' clinical thinking ability. But the role of PBL teaching in improving students' medical knowledge level is not obvious.
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<p><b>OBJECTIVE</b>To develop a real-time PCR assay for the rapid identification of Clostridium(C.)difficile and its toxin.</p><p><b>METHODS</b>TaqMan real-time PCR was developed for the rapid identification of species specific gene(tpi) of C. difficile strains and the toxins A(TcdA), B(TcdB) and truncated toxin A(TcdAT). Sensitivity, specificity and anti-interference ability of these methods were estimated, as well. Feces sampled from fifty diarrhea patients were tested by real-time PCR and compared to the results from VIDAS assay.</p><p><b>RESULTS</b>The detection limits of tpi were 6×10⁻² CFU/µl and 6 × 10⁻¹ CFU/µl in the non-oxin producing and toxin producing strains, respectively. The coefficients of variability(CV) of intra-assay and inter-assay for the detection limits of tpi in the non-toxin producing strain were 2.1% and 2.3% . The CVs of intra-assay and inter-assay for the detection limit of tpi, tcdA, tcdB and tcdAT in the toxin producing strain were 3.0% and 3.4%, 2.9% and 3.2%, 5.3% and 5.7%, 2.7% and 2.8%, respectively. No interference was detected from other genus or species in clostridium. From 50 clinical samples, thirty-nine of them were negative and six of them were positive under the TaqMan-MGB probe technique in accordance with VIDAS. Five samples appeared positive using the TaqMan-MGB probe technique, in which 3 were dubious and 2 were negative under VIDAS.</p><p><b>CONCLUSION</b>The newly developed method was a sensitive and reliable assay for rapid identification of C. difficile and its toxin. This method could be used to screen C. difficile isolates harboring truncated toxin A to avoid misdiagnosis, clinically.</p>
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Proteínas Bacterianas , Toxinas Bacterianas , Clostridioides difficile , Enterotoxinas , Reacción en Cadena en Tiempo Real de la Polimerasa , MétodosRESUMEN
Objective To develop a real-time PCR assay for the rapid identification of Clostridium(C.)difficile and its toxin. Methods TaqMan real-time PCR was developed for the rapid identification of species specific gene(tpi)of C. difficile strains and the toxins A(TcdA),B(TcdB) and truncated toxin A(TcdAT). Sensitivity,specificity and anti-interference ability of these methods were estimated,as well. Feces sampled from fifty diarrhea patients were tested by real-time PCR and compared to the results from VIDAS assay. Results The detection limits of tpi were 6×10-2 CFU/μl and 6 × 10-1 CFU/μl in the non-oxin producing and toxin producing strains,respectively. The coefficients of variability (CV) of intra-assay and inter-assay for the detection limits of tpi in the non-toxin producing strain were 2.1% and 2.3%. The CVs of intra-assay and inter-assay for the detection limit of tpi,tcdA,tcdB and tcdAT in the toxin producing strain were 3.0%and 3.4%,2.9%and 3.2%,5.3%and 5.7%,2.7%and 2.8%,respectively. No interferance was detected from other genus or species in clostridium. From 50 clinical samples,thirty-nine of them were negative and six of them were positive under the TaqMan-MGB probe technique in accordance with VIDAS. Five samples appeared positive using the TaqMan-MGB probe technique,in which 3 were dubious and 2 were negative under VIDAS. Conclusion The newly developed method was a sensitive and reliable assay for rapid identification of C. difficile and its toxin. This method could be used to screen C. difficile isolates harboring truncated toxin A to avoid misdiagnosis,clinically.
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Objective To develop a real-time PCR assay for the rapid identification of Clostridium(C.)difficile and its toxin. Methods TaqMan real-time PCR was developed for the rapid identification of species specific gene(tpi)of C. difficile strains and the toxins A(TcdA),B(TcdB) and truncated toxin A(TcdAT). Sensitivity,specificity and anti-interference ability of these methods were estimated,as well. Feces sampled from fifty diarrhea patients were tested by real-time PCR and compared to the results from VIDAS assay. Results The detection limits of tpi were 6×10-2 CFU/μl and 6 × 10-1 CFU/μl in the non-oxin producing and toxin producing strains,respectively. The coefficients of variability (CV) of intra-assay and inter-assay for the detection limits of tpi in the non-toxin producing strain were 2.1% and 2.3%. The CVs of intra-assay and inter-assay for the detection limit of tpi,tcdA,tcdB and tcdAT in the toxin producing strain were 3.0%and 3.4%,2.9%and 3.2%,5.3%and 5.7%,2.7%and 2.8%,respectively. No interferance was detected from other genus or species in clostridium. From 50 clinical samples,thirty-nine of them were negative and six of them were positive under the TaqMan-MGB probe technique in accordance with VIDAS. Five samples appeared positive using the TaqMan-MGB probe technique,in which 3 were dubious and 2 were negative under VIDAS. Conclusion The newly developed method was a sensitive and reliable assay for rapid identification of C. difficile and its toxin. This method could be used to screen C. difficile isolates harboring truncated toxin A to avoid misdiagnosis,clinically.
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Objective To investigate the changes in the expression of neuromedin U receptor 2 (NMUR2) in spinal dorsal horn in a rat model of bone cancer pain (BCP).Methods Thirty-two female Sprague-Dawley rats,weighing 150-180 g,were randomly divided into 2 groups (n =16 each):sham operation group (group S) and BCP group.BCP was induced by inoculating Walker 256 mammary gland carcinoma cells (1 × 105) into the medullary cavity of left tibia.Heat-killed Walker 256 cells (1 × 105) were injected into the medullary cavity of left tibia in S group.Eight rats were chosen from each group and the paw withdrawal threshold (PWT) to yon Frey filaments was measured at 1 day before operation (baseline) and 1,3,6,9,12 and 15 days after operation.Bone destruction was shown by X-ray at 15 days after operation.At 1 day before operation and 15 days after operation,4 rats in each were chosen and sacrificed,and L4,5 segments of the spinal cord were removed for measurement of the expression of NMUR2 mRNA (by real-time PCR) and protein (using Western blot analysis) in the spinal dorsal horn.Results Compared with S group,the PWT was significantly decreased at day 6-15 after operation and the expression of NMUR2 mRNA and protein in the spinal dorsal horn was up-regulated at 15 days after operation in BCP group (P ＜ 0.05 or 0.01).Compared with the baseline value,the PWT was significantly decreased at day 6-15 after operation and the expression of NMUR2 mRNA and protein in the spinal dorsal horn was up-regulated at 15 days after operation in BCP group (P ＜ 0.05 or 0.01).X-ray showed defect of bone trabecula and cortical bone destruction in BCP group.Conclusion The expression of spinal NMUR2 is up-regulated in rats with BCP and this change may be involved in the development and maintenance of BCP.
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Objective To investigate the effect of dexmedetomidine on bispectral index (BIS) value at loss of consciousness (LOC) caused by propofol given by target-controlled infusion (TCI).Methods One hundred and twenty ASA Ⅰ or Ⅱ patients,aged 20-50 yr,weighing 41-68 kg,scheduled for general surgery were randomly divided into 3 groups (n =40 each):propofol group (group P),dexmedetomidine 0.5 μg/kg + propofol group (group D1P) and dexmedetomidine 1.0 μg/kg + propofol group (group D2P).The patients in each group were randomly assigned into 5 subgroups ( n =8 each):groups P0-4 receiving TCI of propofol with the target effect-site concentration (Ce) set at0,1,2,3 and 4 mg/L respectively.Groups D1P0-4 received iv infusion of dexmedetomidine 0.5 μg/kg at a rate of0.05μg·kg-1 ·min-1 and TCI of propofol with the target Ce set at 0,1,2,3 and 4 mg/L respectively at 5 min after the end of dexmedetomidine infusion.Groups D2 P0-4 received iv infusion of dexmedetomidine 1.0 μg/kg at a rate of 0.1μg· kg- 1· min- 1 and TCI of propofol with the target Ce set at 0,1,2,3 and 4 mg/L respectively at 5 min after the end of dexmedetomidine infusion.Three minutes after TCI of propofol was started,OAA/S score and BIS value were recorded.The OAMS score ≤ 2 was defined as LOC.The EC50 and 95％ confidence interval of propofol for LOC and BIS50 and 95％ confidence interval at LOC were calculated by Probit analysis.Prediction probability (Pk) of BIS value at LOC was calculated using Smith method.Results Compared with group P,EC50 was significantly decreased,BIS50 was significantly increased ( P ＜ 0.05 or 0.01 ),and no significant change was found in Pk in groups D2 P and D1 P ( P ＞ 0.05).There were no significant differences in EC50,BIS50 and Pk between groups D2 P and D1P ( P ＞ 0.05).Conclusion BIS value can accurately predict the level of consciousness during anesthesia with dexmedetomidine and TCI of propofol,but BIS value is increased at LOC.
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Ebola virus (EBOV) causes highly lethal hemorrhagic fever in humans and nonhuman primates and has a significant impact on public health. The nucleoprotein (NP) of EBOV (EBOV-NP) plays a central role in virus replication and has been used as a target molecule for disease diagnosis. In this study, we generated a monoclonal antibody (MAb) against EBOV-NP and mapped the epitope motif required for recognition by the MAb. The MAb generated via immunization of mice with prokaryotically expressed recombinant NP of the Zaire Ebola virus (ZEBOV-NP) was specific to ZEBOV-NP and able to recognize ZEBOV-NP expressed in prokaryotic and eukaryotic cells. The MAb cross-reacted with the NP of the Reston Ebola virus (REBOV), the Cote-d'Ivoire Ebola virus (CIEBOV) and the Bundibugyo Ebola virus (BEBOV) but not with the NP of the Sudan Ebola virus (SEBOV) or the Marburg virus (MARV). The minimal epitope sequence required for recognition by the MAb was the motif PPLESD, which is located between amino acid residues 583 and 588 at the C-terminus of ZEBOV-NP and well conserved among all 16 strains of ZEBOV, CIEBOV and BEBOV deposited in GenBank. The epitope motif is conserved in four out of five strains of REBOV.
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Animales , Ratones , Anticuerpos Monoclonales , Alergia e Inmunología , Ebolavirus , Química , Alergia e Inmunología , Mapeo Epitopo , Métodos , Escherichia coli , Genética , Metabolismo , Ratones Endogámicos BALB C , Nucleoproteínas , Alergia e Inmunología , Proteínas Recombinantes , Genética , Alergia e InmunologíaRESUMEN
ObjectiveTo investigate the effect of dexmedetomidine on the median effectivetarget effectsite concentration (EC50) of remifentanil required for preventing body movement in response to skin incision made under propofol sedation.MethodsForty ASA Ⅰ or Ⅱ patients aged 20-50 yr weighing 45-58 kg scheduled for elective breast tumor excision were randomly allocated into 2 groups ( n =20 each):group remifentanil (group R) and group remifentanil + demedetomidine ( group RD).Sedation was induced with propofol TCI at target plasma concentration of 3.0 mg/L in both groups.In group RD dexmedetomidine 1.0 μg/kg was infused iv over 10 min before start of propofol TCI,while in group R equal volume of normal saline was infused instead of dexmedetomidine.Remifentanil TCI was started with target effect-site concentration set at 3.0 and 2.5 μg/L in groups R and RD respective at 13 min after beginning of propofol TCI.Skin incision (3 cm in length) was made when the target concentrations of propofol and remifentanil TCI were reached.Body movement was assessed by a nurse not involved in this study.EC50 and 95％ confidence interval (CI) of remifentanil were determined by up-and-down technique.The target effect-site concentration was increased or decreased by 20％ depending on the response of the previous patient to skin-incision.ResultsThe EC50 of remifentanil for preventing body movement in response to skin incision performed under propofol sedation was 1.7 μg/L (95％ CI 1.5-1.9 μg/L) and 2.5 μg/L (95％ CI 2.2-2.7μg/L) in groups RD and R respectively.The EC50 of remifentanil was significantly lower in group RD than in group R.ConclusionDexmedetomidine 1.0 μg/kg can decrease EC50 of remifentanil for preventing body movement in response to skin incision made under propofol sedation.
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Aim To observe the effects of promethazine on the analgesia,hypnosis,amnesia and therapeutic index of isoflurane.Methods The experiments were designed to study promethazine on the analgesic effect of isoflurane by hot-plate test and writhing test,and to study the effect of promethazine on the sleeping time of isoflurane by the method of righting reflex,and the amnesia of isoflurane by Morris water maze,and the ED_(50),LD_(50) by sequential method in mice.Results The result of hot-plate test and writhing test indicated that promethazine could enhance the analgesic effect of isoflurane(P＜0.05 or P＜0.01);through the experiment of righting reflex, sleeping time of isoflurane in mice was extended by promethazine(P＜0.01);in Morris water maze experiment, the average latency in the combination of promethazine and isoflurane was longer than that of the promethazine group or isoflurane group(P＜0.05 or P＜0.01), while aiming to the residence time, the combination of the two was shorter than that in the third quadrant(P＜0.01 or P＜0.05),the TCPP of the group of isoflurance was more than that of the combination group;promethazine could decrease the ED_(50) of isoflurance(P＜0.01),but it did not obviously affect its LD_(50)(P＞0.05).Conclusion Promethazine can not only reinforce the effect of isoflurance on analgesia,hypnosis and amnesia, but also boost the therapeutic index of isoflurance.
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Objective To clone,express and characterize a tegument protein gene of Schistosoma japonicum(Sj29),and investigate the immune protection of the recombinant protein against S.japonicum in mice.Methods The gene coding for Sj29 protein was amplified by PCR,and the sequence was analyzed by bioinformatics tools.Partial fragment of Sj29 gene was subcloned into the prokaryotic expression vector pET28c(+).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced the recombinant with IPTG.The recombinant protein(rSj29) was purified by His-binding-resin affinity chromatography and characterized by Western blotting.Three groups each with 10 BALB/c mice were immunized subcutaneously three times(two weeks interval) respectively with 100 ?l recombinant rSj29(0.1 mg/ml),adjuvant or PBS.At the 15th day after the final inoculation,each mouse was challenged by 40 ?2 cercariae of S.japonicum.At the 53th day after infection,the mice were sacrificed to obtain the number of adult worms,number of eggs in liver and feces.Serum samples were collected at pre-immunization and certain time after immuniza-tion,and were analyzed for IgG by ELISA.The localization of rSj29 in worms of different developmental stages was demonstrated by immunofluorescent technique.mRNA expression level of Sj29 gene in worms of different developmental stages and three groups after infection was detected by quantitative real-time PCR.Results A 576 bp Sj29 gene fragment was obtained.The recombinant protein rSj29 with Mr 22 900 was expressed in the form of inclusion body.The recombinant rSj29 can be recognized by sera of mice immunized with rSj29 and sera of infected mice.The number of adult worms(15.4?5.9),number of hepatic eggs(40 143.3?2 995.9) and number of fecal eggs(3 803.9?110.9) in re-combinant protein group were significantly higher than those of PBS control group(20?3.4,49 318.1?6 648.3,5 238.1? 303.5,respectively)(P
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Cellular immunity plays an important role in defense against diseases, such as pathogenic infection,autoimmunity and tumor. With the progress of molecular immunology, mechanisms of T cellular immunity, and the T cell epitopes and functional genomics, studies on the prediction based on data-drived for T cell epitopes has been highlighted, and could be one of the useful tools for application in vaccine development. This review summarizes theory and methodology of prediction for helper T cell epitopes, and their application in vaccine development against parasites, and new research directions are also discussed.