To construct an HSV-1 vector vaccine carrying HIV-1 antigens, HIV-1 gp160, gag, protease and the expression elements were chained together, and then inserted into the internal inverted repeat sequence region of HSV-1 by bacterial artificial chromosome technology. Firstly, HIV-1 gp160 (including type B and C), gag and protease genes were cloned into pcDNA3 in series to generate the pcDNA/gBgp and pcDNA/gCgp, then the recombinant plasmids were transfected into 293FT cells, and HIV-1 antigen was detected from transfected cells by Western blotting. Then the expression cassettes from pcDNA/gBgp and pcDNA/gCgp, comprising HIV-1 antigen genes and expression elements, were cloned into pKO5/BN to generate the shuttle plasmids pKO5/BN/gBgp and pKO5/BN/gCgp. The shuttle plasmids were electroporated into E. coli cells that harbor an HSV-BAC, the recombinant bacteria were screened, and the recombinant DNA was extracted and transfected into Vero cells. The recombinant virus was purified through picking plaques, the virus' DNAs were identified by Southern blotting; HIV-1 antigen was detected from the recombinant HSV-1 infected cells by Western blotting, and the virus' replication competent was analyzed. As the results, gp160 and gag proteins were detected from 293FT cells transfected with pcDNA/gBgp and pcDNA/gCgp by Western blotting. The recombinant bacteria were generated from the E. coli electroporated with pKO5/BN/gBgp or pKO5/BN/gCgp. The recombinant HSV was purified from the Vero cells transfected with the recombinant DNA, the unique DNA fragment was detected from the genome of recombination HSV by Southern blotting; gp120 and gp41 were detected from the infected cells by Western blotting, and the recombinant HSV retained replication competent in mammalian cells. The results indicate that the recombinant HSV carrying HIV-1 gp160, gag and protease genes was generated, the virus retains replication competent in mammalian cells, and could be used as a replicated viral vector vaccine.
Animals ; Cercopithecus aethiops ; Chromosomes, Artificial, Bacterial ; DNA, Recombinant ; genetics ; DNA, Viral ; genetics ; Escherichia coli ; HIV Antigens ; genetics ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; HIV Protease ; genetics ; immunology ; Herpes Simplex Virus Vaccines ; immunology ; Herpesvirus 1, Human ; physiology ; Plasmids ; Transfection ; Vero Cells ; Virus Replication ; gag Gene Products, Human Immunodeficiency Virus ; genetics ; immunology

Objective To explore the application value of noninvasive positive pressure ventilation for acute left heart failure. Methods 86 patients with acute left heart failure were selected and randomly divided into study group and control group,43 patients in each group. Two groups of patients after diagnosis,the control group were treated with conventional treatment, while patients in the study group were given breathing machine with noninvasive positive pressure ventilation treatment. Results Compared with control group,the efficiency and total effective rate in the study group were significantly increased, but the ineffictive rate significantly lower, there were significant differences (P<0. 05). At the same time,patients in the study group,HR and RR were significantly decreased,while PaO2 and SaO2 treatment were significantly higher, there were significant differences (P < 0. 05). Conclusion Use of noninvasive positive pressure ventilation for acute left heart failure,its effects were positive,significant effect on the treatment of acute left heart failure is of great clinical value.

Objective To observe the treatment of patients with limb fractures after taking the bone 7% + bone peptide injection program after treatment. Methods 200 patients with limb fractures in our hospital from January 12, 2016 to May 12, 2017 were selected,and randomly divided into observation group and control group,100 cases in each groups. control group was given bone peptide injection, and observation group was re-used with 7 cm slice, and the healing time and treatment effect of each group were compared. Results The cure time of humeral shaft fractures in patients with limb fractures was (7.59±1.59) w, and the average time (10.97±2.41) w of tibial shaft fractures was (11.15±2.16) w.The total effective rate data 90.00%, both showed better than the control group(P<0.05). Conclusion The combination of bone peptide injection and bone septal syndrome is a method to treat patients with limb fractures, which can effectively shorten the cure time and achieve better therapeutic effect.

Objective: To observe the effect of tripterygium polyglycosid(TP,雷公藤) on airway inflammation and remodeling in rats with asthma.Methods: Forty rats were randomly divided evenly into negative control group,positive control group,normal dose TP group(TPⅠ group) and small dose TP group(TPⅡgroup).The experimental model was induced by ovalbumin sensitization.Number of cells in bronchoalveolar lavage fluid(BALF) was observed by immunocytochemical staining.The expressions of nuclear factor-?B(NF-?B),matrix metalloproteinases-9(MMP-9) and tissue of inhibitor of metalloproteinases-1(TIMP-1) were detected by immunohistochemistry method.Results: Compared with the negative control group,the counts of lymphocyte,neutrophilic leukocyte,macrophage and eosinocyte in BALF were elevated significantly,positive cell percentages of NF-?B, MMP-9 and TIMP-1 in lung tissues increased greatly in positive control group,the differences being significant(all P

ObjectiveInvestigate the value and feasibility of image registration with reverse rotation in lateral cerebral DSA.Methods ( 1 ) Experimental study:the target images were subtracted directly,and subtracted again after reverse rotation.Software of registration and subtraction with reverse rotation edited by the author utilizing Visual Basic.The function of the automatic angle detection by the software were evaluated to see whether it detected the angle of line.The subtraction function of DSA by the software was evaluated.(2) Clinical retrospective study:the untreated mask and target inages of 15 patients with motion along vertical axis during lateral cerebral DSA were uploaded to the software.The target images were processed with and without the software to get two sets of images.( 3 ) Evaluation:four experienced radiologists read and compared the two sets of the images,and graded their findings.Results ( 1 ) The automatic detection by the software suggested that the target images should be rotated counterclockwise 1.3°. The subtraction result of the software was satisfactory.(2)In the 15 sets of images,there were only three sets of images deemed optimal after traditional subtraction.After reverse rotation,artifacts were significantly reduced and the image sharper. There were ten cases with significant artifacts after traditional subtraction,and those images were sharper and showed more peripheral vessels after reverse rotation. The traditional subtraction images of two sets could not be interpreted,the reverse rotation registration images reached the diagnostic quality.(3)Subjective evaluation: there were more information and less noise and distortion in the registration images with reverse rotation than in the traditional subtraction. But the image resolution decreased slightly after reverse rotation registration.ConclusionThe registration of digital angiography with reverse rotation can improve the image quality in lateral cerebral DSA.

Eighty one infants with severe asthma attacks were randomly divided into three groups:budesonide group (budesonide suspension + ventolin inhalation),methylprednisolone group (Ventolin inhalation + intravenous methylprednisoloue) and ventolin group (ventolin inhalation alone).Compared with the pre-treatment,the respiratory rate,heart rate,wheeze score,self-feeling score of three groups were gradually reduced (q=2.96-163.37,P＜0.05 or 0.01).The respiratory rate,heart rate of ventolin group was significantly higher than those of budesonide group (q=3.08,4.10,P＜0.05) and methylprednisolone group (q=3.24,3.34,P＜0.05) 4 h after treatment,wheeze score,self-feeling score of ventolin group was significantly higher than budesonide group (q=5.63-23.63,P＜0.01) and methylprednisolone group (q=6.76-23.72,P＜0.01) 4 and 12 h after treatment.Results indicate that budesonide suspension can achieve the same effect as intravenous methylprednisolone and bronchodilators alone may not effectively control the severe asthma attack in infants.

Objective To explore the clinical efficacy of Qingyi decoction (a preparation of Chinese herbal medicine) combined with DAR (combined use of dexamethasone,anisodamine and rhubarb) for the treatment of acute pancreatitis.Methods A total of 387 eligible patients met the criteria of acute pancreatitis were enrolled from January 2005 to April 2012 for prospective study.All patients,mild acute pancreatitis (MAP) or severe acute pancreatitis (SAP),were divided (random niumber) into four groups,namely conventional therapy (T),DAR therapy (DAR),Qingyi decoction therapy (Q) and Qingyi decoction combined with DAR therapy (Q + DAR).Outcome,fasting time,serum amylase,abdominal pain relief time,pancreatic or peri-pancreatic complications and average hospital-stay were analvzed with SPSS 13.0 statistic software.P ＜ 0.05 was considered statistically significant.Results None of MAP patients died.Of SAP patients,there was no difference in mortality among different groups (P ＞ 0.05).length of fasting time,tine elapsed for abdominal pain relief,time required for normalized serum amylase level and length of hospital stay in MAP patients were significantly shorter than those in SAP patients regardless of different therapies (P ＜ 0.05).Either patients of SAP or MAP treated with Q + DAR or DAR suffered shorter length of time than those treated with T or Q in respect of fasting,abdominal paiu relief,serum amylase level normalization and hospital stay (P ＜ 0.05).For SAP or MAP patients,there was no difference in abdominal pain relief time between receiving DAR and Q + DAR treatment (P ＞ 0.05),but the fasting time in Q + DAR was shorter than that in DAR (P ＜ 0.05).Patients with SAP were more likely to suffer pancreatic or per-pancreatic complications than those with MAP,but there was no difference for SAP or MAP with different treatments.Conclusions DAR or Q + DAR was an alternative to conventional treatment for MAP or SAP,and they were both superior to conventional treatment.And Q + DAR was more advantageous than DAR when fasting time,hospital-stay time and cost were considered.

Objective To investigate the first medical contact to balloon ( FMC2B) time in our center and to identify the influencing factors .Methods This is a retrospective study conducted in the heart center of Beijing Luhe Hospital . A total of 140 patients undergoing primary percutaneous coronary intervention ( PCI) were enrolled between July 2013 to September 2014.Demographic data , clinical risk factors and the emergency process were evaluated .All the patients were categorized into 2 groups including:the conformed group ( patients with FMC2B<120 min for non-PCI-capable hospital and <90 min for direct arrival at Luhe hospital, n=59) and the unconformed group (n=81).Multivariant regression aralysis was done to analyse factors influencing FMC 2B time.Results Among the enrolled 140 patients, 58 patients were initially seen in a non-PCI-capable hospital , 31 patients were directly sent to Luhe hospital by ambulance and 51 patients arrived by themselves.The median FMC2B time was 106.16 min (interquartile range [ IQR ]: 77.37 -165.52 min ) and 42.1% ( 59/140 ) of the patients achieved the current recommended FMC2B time.In a multivariate logistic analysis , FMC to electrocardiographic ( ECG) within 10 min ( OR=5.61 , 95% CI 1.91-16.88 ) , admission during normal working hours ( OR=5.11 , 95%CI 1.88-13.85 ) , patient′s education level of high school or above ( OR=4.16 , 95%CI 1.53-11.34 ) , awareness of heart diseases ( OR =2.58, 95% CI 1.13 -5.91 ) were predictors of improving FMC2B. Transfer for primary PCI (OR=0.37, 95% CI 0.15-0.92) increased FMC2B.Conclusions Less than half of the patients with primary PCI achieved the goal of guidelines′recommended FMC2B time.Initial ECG, admission during normal working hours , patient′s education level and awareness of heart diseases and transfer for primary PCI are the independent predictors of FMC 2B time.

Objective:To understand the current work situation of clinical pharmacy in medical institutions in Wuhan city .Meth-ods:A questionnaire survey was employed to investigate the stuffing situation of clinical pharmacists , quality of clinical pharmacists and clinical pharmacy development in medical institutions above class II in Wuhan city .The results were analyzed statistically .Re-sults:There was notable difference in the situation of clinical pharmacist among medical institutions above class Ⅱin Wuhan city , and so was in the development of clinical pharmacy .Conclusion:Clinical pharmacy work in medical institutions still demands much atten-tion and support from the relevant departments , and clinical pharmacists own need make great efforts constantly .

Objective To compare the sensitivity and specificity of dot immunogold method (DIM) and particle agglutination (PA) for the diagnosis of mycoplasma pneumoniae (MP) infection. Methods The 190 serum specimens of 113 children with mycoplasmal pneumonia (infection group) and 50 serum specimens of 50 health children (health group) were tested for MP by PA and DIM- A and B. Results In infection group, the positive rates of DIM- A and B were 82.63% (157/190) and 84.74%(161/190), and there was no statistical difference (χ2 = 0.31, P>0.05); the positive rate of PA (titer ≥1:160) was 70.00%(133/190), the positive rate of PA was significantly lower than that in DIM-A and B, and there were statistical differences (P<0.05). In infection group, with the increase of PA titer, the positive rate of DIM was gradually increased, and there was a correlation between 2 methods (rA=0.972, rB=0.830);the positive rates of DIM- A and B in serum specimens of PA negative were 40.62%(13/32) and 53.12%(17/32), and in the PA titer of 1:5120, there was still a negative result in DIM. In health group, the positive rates of DIM-A and B were 26.00% (13/50) and 28.00% (14/50), and there was no statistical difference (χ2 = 0.66, P>0.05); the positive of PA was 8.00% (4/50), the positive rate of PA was significantly lower than that in DIM- A and B, and there were statistical differences (P<0.05 or<0.01). Conclusions Compared with the PA, DIM has low sensitivity and poor specificity for clinical diagnosis. DIM is not suitable for clinical diagnosis of MP infection.